Methods and devices for sample collection and sample separation

ABSTRACT

Methods and devices are provided for sample collection and sample separation. In one embodiment, a device is provided for use with a formed component liquid sample, the device comprising at least one sample inlet for receiving said sample; at least a first outlet for outputting only a liquid portion of the formed component liquid sample; at least a second outlet for outputting the formed component liquid sample at least a first material mixed therein.

BACKGROUND

A blood sample for use in laboratory testing is often obtained by way ofvenipuncture, which typically involves inserting a hypodermic needleinto a vein on the subject. Blood extracted by the hypodermic needle maybe drawn directly into a syringe or into one or more sealed vials forsubsequent processing. When a venipuncture may be difficult orimpractical such as on a newborn infant, a non-venous puncture such as aheel stick or other alternate site puncture may be used to extract ablood sample for testing. After the blood sample is collected, theextracted sample is typically packaged and transferred to a processingcenter for analysis.

Unfortunately, conventional sample collection and testing techniques ofbodily fluid samples have drawbacks. For instance, except for the mostbasic tests, blood tests that are currently available typically requirea substantially high volume of blood to be extracted from the subject.Because of the high volume of blood, extraction of blood from alternatesample sites on a subject, which may be less painful and/or lessinvasive, are often disfavored as they do not yield the blood volumesneeded for conventional testing methodologies. In some cases, patientapprehension associated with venipuncture may reduce patient compliancewith testing protocol. Furthermore, the traditional collection techniqueadds unnecessary complexity when trying to separate a single bloodsample into different containers for different pre-analyticalprocessing.

SUMMARY

At least some of the disadvantages associated with the prior art areovercome by one or more embodiments of the devices, systems, or methodsdescribed herein.

In one embodiment, a device is provided for use with a formed componentliquid sample, the device comprising at least one sample inlet forreceiving said sample; at least a first outlet for outputting only aliquid portion of the formed component liquid sample; at least a secondoutlet for outputting the formed component liquid sample at least afirst material mixed therein.

It should be understood that one or more of the following features maybe adapted for use with any of the embodiments described herein. By wayof non-limiting example, the body may a first pathway fluidicallycouples the sample inlet with the first outlet. Optionally, a secondpathway fluidically couples the sample inlet with the second outlet.Optionally, a separation material along the first pathway configured toremove said formed component from the sample prior to outputting at thefirst outlet. Optionally, the separation material and a distributor areconfigured to have an interface that provides a multi-mode samplepropagation pattern wherein at least a first portion is propagatinglaterally within the separator and a second portion is propagatingthrough the channels of the distributor over the separator. Optionally,there is at least 50 mm² surface area of separator per 30 uL of sampleto filter. Optionally, there is at least 60 mm² surface area ofseparator per 30 uL of sample to filter. Optionally, there is at least70 mm² surface area of separator per 30 uL of sample to filter.Optionally, the inlet directs the sample to primarily contact a planarportion of separator surface, and not a lateral edge of the separator.Optionally, the amount of time for sample to fill the first pathway andreach the first outlet is substantially equal to the time for sample tofill the second pathway and reach the second outlet. Optionally, thefirst pathway comprises a portion configured in a distributed pattern ofchannels over the filtration material to preferentially direct thesample over the surface of the separation material in a pre-determinedconfiguration. Optionally, at least a portion of the separation materialis coupled to a vent which contacts the membrane in a manner that thevent is only accessible fluidically by passing through the separationmaterial. Optionally, containers have interiors under vacuum pressurethat draw sample therein. Optionally, the separation material is held inthe device under compression. Optionally, the separation materialcomprises an asymmetric porous membrane. Optionally, the separationmaterial is a mesh. Optionally, the separation material comprisespolyethylene (coated by ethylene vinyl alcohol copolymer). Optionally,at least a portion of the separation material comprises apolyethersulfone. Optionally, at least a portion of the separationmaterial comprises an asymmetric polyethersulfone. Optionally, at leasta portion of the separation material comprises polyarylethersulfone.Optionally, at least a portion of the separation material comprises anasymmetric polyarylethersulfone. Optionally, at least a portion of theseparation material comprises a polysulfone. Optionally, the separationmaterial comprises an asymmetric polysulfone. Optionally, the separationmaterial comprises a cellulose or cellulose derivative material.Optionally, the separation material comprises polypropylene (PP).Optionally, the separation material comprises polymethylmethacrylate(PMMA). In one non-limiting example, the separation material comprises apolymer membrane wherein filtrate exit surface of the membrane comprisesa relatively open pore structure and the opposite surface comprises amore open pore structure and wherein the supporting structure comprisesasymmetry through at least 50% of the supporting structure but no morethan 95% of the supporting structure, the membrane having surface poresat the minimum surface of a mean diameter of at least about 1 micron andhaving a flow rate of greater than about 4 cm/min/psi. Optionally, theflow rate of the material, unassisted, is between 1 cm/min/psi and 4.3 2cm/min/psi.

In one embodiment described herein, a device for collecting a bodilyfluid sample from a subject is provided comprising: at least two samplecollection pathways configured to draw the bodily fluid sample into thedevice from a single end of the device in contact with the subject,thereby separating the fluid sample into two separate samples; a secondportion comprising a plurality of sample containers for receiving thebodily fluid sample collected in the sample collection pathways, thesample containers operably engagable to be in fluid communication withthe sample collection pathways, whereupon when fluid communication isestablished, the containers provide a motive force to move a majority ofthe two separate samples from the pathways into the containers.Optionally, the device includes a separation material along one of thesample collection pathways, the material configured to remove formedcomponents from the sample when outputting to at least one of the samplecontainers.

In another embodiment described herein, a device for collecting a bodilyfluid sample is provided comprising: a first portion comprising at leastone fluid collection location leading to at least two sample collectionpathways configured to draw the fluid sample therein via a first type ofmotive force; a second portion comprising a plurality of samplecontainers for receiving the bodily fluid sample collected in the samplecollection pathways, the sample containers operably engagable to be influid communication with the sample collection pathways, whereupon whenfluid communication is established, the containers provide a secondmotive force different from the first motive force to move a majority ofthe bodily fluid sample from the pathways into the containers; whereinat least one of the sample collection pathways comprises a fillindicator to indicate when a minimum fill level has been reached andthat at least one of the sample containers can be engaged to be in fluidcommunication with at least one of the sample collection pathways.Optionally, the device includes a separation material along one of thesample collection pathways, the material configured to remove formedcomponents from the sample when outputting to at least one of the samplecontainers.

In another embodiment described herein, a device for collecting a bodilyfluid sample is provided comprising a first portion comprising at leasttwo sample collection channels configured to draw the fluid sample intothe sample collection channels via a first type of motive force, whereinone of the sample collection channels has an interior coating designedto mix with the fluid sample and another of the sample collectionchannels has another interior coating chemically different from saidinterior coating; a second portion comprising a plurality of samplecontainers for receiving the bodily fluid sample collected in the samplecollection channels, the sample containers operably engagable to be influid communication with the collection channels, whereupon when fluidcommunication is established, the containers provide a second motiveforce different from the first motive force to move a majority of thebodily fluid sample from the channels into the containers; whereincontainers are arranged such that mixing of the fluid sample between thecontainers does not occur. Optionally, the device includes a separatoralong one of the sample collection channels, the separator configured toremove formed component from the sample when outputting to at least oneof the sample containers.

In another embodiment described herein, a device for collecting a bodilyfluid sample is provided comprising: a first portion comprising aplurality of sample collection channels, wherein at least two of thechannels are configured to simultaneously draw the fluid sample intoeach of the at least two sample collection channels via a first type ofmotive force; a second portion comprising a plurality of samplecontainers for receiving the bodily fluid sample collected in the samplecollection channels, wherein the sample containers have a firstcondition where the sample containers are not in fluid communicationwith the sample collection channels, and a second condition where thesample containers are operably engagable to be in fluid communicationwith the collection channels, whereupon when fluid communication isestablished, the containers provide a second motive force different fromthe first motive force to move bodily fluid sample from the channelsinto the containers. Optionally, the device includes a separator alongone of the sample collection channels, the separator configured toremove formed component from the sample when outputting to at least oneof the sample containers.

In another embodiment described herein, a sample collection device isprovided comprising: (a) a collection channel comprising a first openingand a second opening, and being configured to draw a bodily fluid samplevia capillary action from the first opening towards the second opening;and (b) a sample container for receiving the bodily fluid sample, thecontainer being engagable with the collection channel, having aninterior with a vacuum therein, and having a cap configured to receive achannel; wherein the second opening is defined by a portion thecollection channel configured to penetrate the cap of the samplecontainer, and to provide a fluid flow path between the collectionchannel and the sample container, and the sample container has aninterior volume no greater than ten times larger than the interiorvolume of the collection channel. Optionally, the device comprises aseparator along one of the sample collection channel, the separatorconfigured to remove formed component from the sample prior to and whenoutputting to the sample container.

In another embodiment described herein, a sample collection device isprovided comprising: (a) a collection channel comprising a first openingand a second opening, and being configured to draw a bodily fluid samplevia capillary action from the first opening towards the second opening;(b) a sample container for receiving the bodily fluid sample, thecontainer being engagable with the collection channel, having aninterior with a vacuum therein, and having a cap configured to receive achannel; and (c) an adaptor channel configured to provide a fluid flowpath between the collection channel and the sample container, having afirst opening and a second opening, the first opening being configuredto contact the second opening of the collection channel, the secondopening being configured to penetrate the cap of the sample container.Optionally, the device comprises a separator along one of the samplecollection channel, the separator configured to remove formed componentfrom the sample prior to and when outputting to the sample container.

In another embodiment described herein, a sample collection device isprovided comprising: (a) a body, containing a collection channel , thecollection channel comprising a first opening and a second opening, andbeing configured to draw a bodily fluid via capillary action from thefirst opening towards the second opening; (b) a base, containing asample container for receiving the bodily fluid sample, the samplecontainer being engagable with the collection channel, having aninterior with a vacuum therein, and having a cap configured to receive achannel; and (c) a support, wherein, the body and the base are connectedto opposite ends of the support, and are configured to be movablerelative to each other, such that sample collection device is configuredto have an extended state and a compressed state, wherein at least aportion of the base is closer to the body in the extended state of thedevice than in the compressed state, the second opening of thecollection channel is configured to penetrate the cap of the samplecontainer, in the extended state of the device, the second opening ofthe collection channel is not in contact with the interior of the samplecontainer, and in the compressed state of the device, the second openingof the collection channel extends into the interior of the samplecontainer through the cap of the container, thereby providing fluidiccommunication between the collection channel and the sample container.Optionally, the device comprises a separator along one of the samplecollection channel, the separator configured to remove formed componentfrom the sample prior to and when outputting to the sample container.

In another embodiment described herein, a sample collection device isprovided comprising: (a) a body, containing a collection channel , thecollection channel comprising a first opening and a second opening, andbeing configured to draw a bodily fluid via capillary action from thefirst opening towards the second opening; (b) a base, containing asample container for receiving the bodily fluid sample, the samplecontainer being engagable with the collection channel, having aninterior with a vacuum therein and having a cap configured to receive achannel; (c) a support, and (d) an adaptor channel, having a firstopening and a second opening, the first opening being configured tocontact the second opening of the collection channel, and the secondopening being configured to penetrate the cap of the sample container,wherein, the body and the base are connected to opposite ends of thesupport, and are configured to be movable relative to each other, suchthat sample collection device is configured to have an extended stateand a compressed state, wherein at least a portion of the base is closerto the body in the extended state of the device than in the compressedstate, in the extended state of the device, the adaptor channel is notin contact with one or both of the collection channel and the interiorof the sample container, and in the compressed state of the device, thefirst opening of the adaptor channel is in contact with the secondopening of the collection channel, and the second opening of the adaptorchannel extends into the interior of the sample container through thecap of the container, thereby providing fluidic communication betweenthe collection channel and the sample container. Optionally, the devicecomprises a separator along one of the sample collection channel, theseparator configured to remove formed component from the sample prior toand when outputting to the sample container.

In another embodiment described herein, a device for collecting a fluidsample from a subject is provided comprising: (a) a body containing acollection channel, the collection channel comprising a first openingand a second opening, and being configured to draw a bodily fluid viacapillary action from the first opening towards the second opening; (b)a base, engagable with the body, wherein the base supports a samplecontainer, the container being engagable with the collection channel,having an interior with a vacuum therein, and having a cap configured toreceive a channel; wherein the second opening of the collection channelis configured to penetrate the cap of the sample container, and toprovide a fluid flow path between the collection channel and the samplecontainer. Optionally, the device comprises a separator along one of thesample collection channel, the separator configured to remove formedcomponent from the sample prior to and when outputting to the samplecontainer.

In another embodiment described herein, a device for collecting a fluidsample from a subject is provided comprising: (a) a body containing acollection channel, the collection channel comprising a first openingand a second opening, and being configured to draw a bodily fluid viacapillary action from the first opening towards the second opening; (b)a base, engagable with the body, wherein the base supports a samplecontainer, the sample container being engagable with the collectionchannel, having an interior with a vacuum therein and having a capconfigured to receive a channel; and (c) an adaptor channel, having afirst opening and a second opening, the first opening being configuredto contact the second opening of the collection channel, and the secondopening being configured to penetrate the cap of the sample container.Optionally, the device comprises a separator along one of the samplecollection channel, the separator configured to remove formed componentfrom the sample prior to and when outputting to the sample container.

It should be understood that one or more of the following features maybe adapted for use with any of the embodiments described herein. By wayof non-limiting example, the body may comprise of two collectionchannels. Optionally, the interior of the collection channel(s) arecoated with an anticoagulant. Optionally, the body comprises a firstcollection channel and a second collection channel, and the interior ofthe first collection channel is coated with a different anticoagulantthan the interior of the second collection channel. Optionally, thefirst anticoagulant is ethylenediaminetetraacetic acid (EDTA) and thesecond anticoagulant is different from EDTA. Optionally, the firstanticoagulant is citrate and the second anticoagulant is different fromcitrate. Optionally, the first anticoagulant is heparin and the secondanticoagulant is different from heparin. Optionally, one anticoagulantis heparin and the second anticoagulant is EDTA. Optionally, oneanticoagulant is heparin and the second anticoagulant is citrate.Optionally, one anticoagulant is citrate and the second anticoagulant isEDTA. Optionally, the body is formed from an optically transmissivematerial. Optionally, the device includes the same number of samplecontainers as collection channels. Optionally, the device includes thesame number of adaptor channels as collection channels. Optionally, thebase contains an optical indicator that provides a visual indication ofwhether the sample has reached the sample container in the base.Optionally, the base is a window that allows a user to see the containerin the base. Optionally, the support comprises a spring, and springexerts a force so that the device is at the extended state when thedevice is at its natural state. Optionally, the second opening of thecollection channel or the adaptor channel is capped by a sleeve, whereinsaid sleeve does not prevent movement of bodily fluid via capillaryaction from the first opening towards the second opening. Optionally,the sleeve contains a vent. Optionally, each collection channel can holda volume of no greater than 500 uL. Optionally, each collection channelcan hold a volume of no greater than 200 uL. Optionally, each collectionchannel can hold a volume of no greater than 100 uL. Optionally, eachcollection channel can hold a volume of no greater than 70 uL.Optionally, each collection channel can hold a volume of no greater than500 uL. Optionally, each collection channel can hold a volume of nogreater than 30 uL. Optionally, the internal circumferential perimeterof a cross-section of each collection channel is no greater than 16 nmi.Optionally, the internal circumferential perimeter of a cross-section ofeach collection channel is no greater than 8 mm. Optionally, theinternal circumferential perimeter of a cross-section of each collectionchannel is no greater than 4 mm. Optionally, the internalcircumferential perimeter is a circumference. Optionally, the devicecomprises a first and a second collection channel, and the opening ofthe first channel is adjacent to an opening of said second channel, andthe openings are configured to draw blood simultaneously from a singledrop of blood. Optionally, the opening of the first channel and theopening of the second channel have a center-to-center spacing of lessthan or equal to about 5 mm. Optionally, each sample container has aninterior volume no greater than twenty times larger than the interiorvolume of the collection channel with which it is engagable. Optionally,each sample container has an interior volume no greater than ten timeslarger than the interior volume of the collection channel with which itis engagable. Optionally, each sample container has an interior volumeno greater than five times larger than the interior volume of thecollection channel with which it is engagable. Optionally, each samplecontainer has an interior volume no greater than two times larger thanthe interior volume of the collection channel with which it isengagable. Optionally, establishment of fluidic communication betweenthe collection channel and the sample container results in transfer ofat least 90% of the bodily fluid sample in the collection channel intothe sample container.

It should be understood that one or more of the following features maybe adapted for use with any of the embodiments described herein.Optionally, establishment of fluidic communication between thecollection channel and the sample container results in transfer of atleast 95% of the bodily fluid sample in the collection channel into thesample container. Optionally, establishment of fluidic communicationbetween of the collection channel and the sample container results intransfer of at least 98% of the bodily fluid sample in the collectionchannel into the sample container. Optionally, establishment of fluidiccommunication between the collection channel and the sample containerresults in transfer of the bodily fluid sample into the sample containerand in no more than ten uL of bodily fluid sample remaining in thecollection channel. Optionally, establishment of fluidic communicationbetween the collection channel and the sample container results intransfer of the bodily fluid sample into the sample container and in nomore than five uL of bodily fluid sample remaining in the collectionchannel. Optionally, engagement of the collection channel with thesample container results in transfer of the bodily fluid sample into thesample container and in no more than 2 uL of bodily fluid sampleremaining in the collection channel. Optionally, the channels have across-sectional shape characterized by a greater width than height.Optionally, the channels are distributed in a pattern where at leastsome of the channels intersect other channels to form a grid pattern.

Optionally, the sample inlet introduces the sample over the manifold.Optionally, the sample inlet introduces the sample along at least anedge or lateral side portion of the manifold. Optionally, the sampleinlet introduces the sample over the manifold and at least a lateralside portion of the manifold.

In another embodiment described herein, a method is provided comprisingcontacting one end of a sample collection device to a bodily fluidsample to split the sample into at least two portions by drawing thesample into at least two collection channels of the sample collectiondevice by way of a first type of motive force; establishing fluidcommunication between the sample collection channels and the samplecontainers after a desired amount of sample fluid has been confirmed tobe in at least one of the collection channels, whereupon the containersprovide a second motive force different from the first motive force tomove each of the portions of bodily fluid sample into their respectivecontainers.

In another embodiment described herein, a method is provided comprisingmetering a minimum amount of sample into at least two channels by usinga sample collection device with at least two of the sample collectionchannels configured to simultaneously draw the fluid sample into each ofthe at least two sample collection channels via a first type of motiveforce; after a desired amount of sample fluid has been confirmed to bein the collection channels, fluid communication is established betweenthe sample collection channels and the sample containers, whereupon thecontainers provide a second motive force different from the first motiveforce use to collect the samples to move bodily fluid sample from thechannels into the containers.

In another embodiment described herein, a method of collecting a bodilyfluid sample is provided comprising (a) contacting a bodily fluid samplewith a device comprising a collection channel, the collection channelcomprising a first opening and a second opening, and being configured todraw a bodily fluid via capillary action from the first opening towardsthe second opening, such that the bodily fluid sample fills thecollection channel from the first opening through the second opening;(b) establishing a fluid flow path between the collection channel andthe interior of a sample container , said sample container having aninterior volume no greater than ten times larger than the interiorvolume of the collection channel and having a vacuum prior toestablishment of the fluid flow path between the collection channel andthe interior of the sample container, such that establishment of thefluid flow path between the collection channel and the interior of thesample container generates a negative pressure at the second opening ofthe collection channel, and the fluidic sample is transferred from thecollection channel to the interior of the sample container.

In another embodiment described herein, a method of collecting a bodilyfluid sample is provided comprising (a) contacting a bodily fluid samplewith any collection device as described herein, such that the bodilyfluid sample fills the collection channel from the first opening throughthe second opening of at least one of the collection channel(s) in thedevice; and (b) establishing a fluid flow path between the collectionchannel and the interior of the sample container , such thatestablishing a fluid flow path between the collection channel and theinterior of the sample container generates a negative pressure at thesecond opening of the collection channel, and the fluidic sample istransferred from the collection channel to the interior of the samplecontainer.

It should be understood that one or more of the following features maybe adapted for use with any of the embodiments described herein.Optionally, the collection channel and the interior of the samplecontainer are not brought into fluid communication until the bodilyfluid reaches the second opening of the collection channel. Optionally,the device comprises two collection channels, and the collectionchannels and the interior of the sample containers are not brought intofluidic communication until the bodily fluid reaches the second openingof both collection channels. Optionally, the second opening of thecollection channel in the device is configured to penetrate the cap ofthe sample container, and wherein a fluidic flow path between the secondopening of the collection channel and the sample container isestablished by providing relative movement between the second opening ofthe collection channel and the sample container, such that the secondopening of the collection channel penetrates the cap of the samplecontainer. Optionally, the device comprises an adaptor channel for eachcollection channel in the device, the adaptor channel having a firstopening and a second opening, the first opening being configured tocontact the second opening of the collection channel, and the secondopening being configured to penetrate the cap of the sample container,and wherein a fluidic flow path between the collection channel and thesample container is established by providing relative movement betweentwo or more of: (a) the second opening of the collection channel, (b)the adaptor channel, and (c) the sample container, such that the secondopening of the adaptor channel penetrates the cap of the samplecontainer.

In another embodiment described herein, a method for collecting a bodilyfluid sample from a subject is provided comprising: (a) bringing adevice comprising a first channel and a second channel into fluidiccommunication with a bodily fluid from the subject, each channel havingan input opening configured for fluidic communication with said bodilyfluid, each channel having an output opening downstream of the inputopening of each channel, and each channel being configured to draw abodily fluid via capillary action from the input opening towards theoutput opening; (b) bringing, through the output opening of each of thefirst channel and the second channel, said first channel and said secondchannel into fluidic communication with a first container and a secondcontainer, respectively; and (c) directing said bodily fluid within eachof said first channel and second channel to each of said first containerand second container with the aid of: (i) negative pressure relative toambient pressure in said first container or said second container,wherein said negative pressure is sufficient to effect flow of saidbodily fluid through said first channel or said second channel into itscorresponding container, or (ii) positive pressure relative to ambientpressure upstream of said first channel or said second channel, whereinsaid positive pressure is sufficient to effect flow of said whole bloodsample through said first channel or said second channel into itscorresponding container.

In another embodiment described herein, a method of manufacturing asample collection device is provided comprising forming one portion of asample collection device having at least two channels configured tosimultaneously draw the fluid sample into each of the at least twosample collection channels via a first type of motive force; formingsample containers, whereupon the containers are configured to be coupledto the sample collection device to the provide a second motive forcedifferent from the first motive force use to collect the samples to movebodily fluid sample from the channels into the containers.

In another embodiment described herein, computer executable instructionsare provided for performing a method comprising: forming one portion ofa sample collection device having at least two channels configured tosimultaneously draw the fluid sample into each of the at least twosample collection channels via a first type of motive force.

In another embodiment described herein, computer executable instructionsfor performing a method comprising: forming sample containers, whereuponthe containers are configured to be coupled to the sample collectiondevice to provide a second motive force different from the first motiveforce use to collect the samples to move bodily fluid sample from thechannels into the containers.

In yet another embodiment described herein, a device for collecting abodily fluid sample from a subject, the device comprising: means fordrawing the bodily fluid sample into the device from a single end of thedevice in contact with the subject, thereby separating the fluid sampleinto two separate samples; means for transferring the fluid sample intoa plurality of sample containers, wherein the containers provide amotive force to move a majority of the two separate samples from thepathways into the containers.

In one embodiment, the desired range of channel surface area relative tothe surface area of the separator on that side of the separator is inthe range of about 35% to 70%. Optionally, the desired range of channelsurface area relative to the surface area of the separator on that sideof the separator is in the range of about 40% to 70%. Optionally, thedesired range of channel surface area relative to the surface area ofthe separator on that side of the separator is in the range of about 50%to 60%.

In yet another embodiment, a method is provided comprising collecting abodily fluid sample into a collection channel, the collection channelcomprising a first opening and a second opening, and being configured todraw the bodily fluid via capillary action from the first openingtowards the second opening; and using a separator along the samplecollection channel to remove formed component from the sample prior toand when outputting to the sample container.

In yet another embodiment, a method is provided comprising collecting abodily fluid sample into device having a first collection channel and asecond collection channel, the collection channel comprising a firstopening and a second opening, and being configured to draw the bodilyfluid via capillary action from the first opening towards the secondopening; using a separator along the first sample collection channel toremove formed component from the sample prior to and when outputting tothe sample container; wherein when the bodily fluid sample is blood, thedevice outputs both blood and plasma, each from separate outlets, fromthe one sample collected into the device.

In one embodiment described herein, it is desirable to use separationmaterials on a bodily fluid to allow for plasma-based assays. Thedesired list of assays includes not only large molecules such asproteins and lipids, but also smaller metabolites such as those that arepart of the complete metabolic panel and examples include but are notlimited to glucose, calcium, magnesium, etc . . . . Since the plasmaseparation materials were not primarily designed for these assays butfor use in select types of test-strip based assays, thehemolysis-preventing agent used in these materials can interfere withother assay chemistries.

In the case of at least some bodily fluid separation materials describedherein, the separation material may have a coating of a protectivematerial such as but not limited to an anti-hemolytic material likesingle and/or double alkyl chain N-oxides of tertiary amines (NTA).Alternatively, separation material coating can constitute a combinationof an anti-hemolytic (such as surfactant, protein, sugar, or acombination of these), alongside an anti-coagulant (such as EDTA and itsderivatives or Heparin). NTA generally does not interfere with severallarge molecule assays. NTA, however, is a chelating agent that stronglybinds to di-valent cations such as calcium and magnesium ions.Unfortunately, this results in a very strong interference in certainassays used to measure, for example and not limitation, calcium andmagnesium concentrations and also for assays where Ca and Mg areco-factors for enzymes which are part of the reaction. This can resultin significant errors for such assays.

One or more of the embodiments described herein provide the benefits ofthe anti-hemolytic material but also provide a much reduced downsideeffect of the anti-hemolytic material leaching into the bodily fluid andaltering the assay results. It should be understood that the coating, insome embodiments, can be one or more of the following: anti-coagulant,anti-hemolytic, and molecules for surface coverage. Any of these mayinterfere with assays. Some embodiments disclosed herein are directedtoward multi-region separation material structures with captureregion(s) and pass-through region(s) with different surface treatments.

Optionally, these separation materials may be asymmetric ornon-asymmetric separation materials. Some embodiments have bi-layer,tri-layer, or other multi-layer configurations. Some embodiments may becontinuously asymmetric with the asymmetric region extending from anupper surface of the material to a lower surface of the material.Optionally, some embodiment may have only one or more portions of thematerial that are asymmetric while one or more other regions areisotropic in terms of pore size. Some embodiments may have an asymmetricmaterial that is then bonded to at least another material that isisotropic to create a desired pore size distribution profile. In such anembodiment, the asymmetric region may have the larger pore sizes and becoated with anti-hemolytic material. Some embodiments can haveseparation materials with gradation in coating material thickness and/orcoverage to position material such as the hemolysis-preventing materialin areas where the material is likely to be in contact with formedcomponents captured by the separation material.

By way of non-limiting example, some separation materials may be washedin a manner the preferentially removes the anti-hemolytic material fromat least one region of the separation material, such as the innerportions of the separation material, but not the exterior portions thatare more likely to come into direct contact with formed components ofthe bodily fluid. Other variations or alternative coating schemes tocreate separation materials or filter structures with areas of leachingand non-leaching materials are not excluded. Optionally, separationmaterials can also be coated with at least two different materials thatmay both leach into the bodily fluid, but at least one of thesematerials that may leach into the fluid does not impact assaymeasurements and can be used to overcoat the other material and thusdecrease the surface area exposure of the other material to the bodilyfluid.

Optionally, the separation material comprises an asymmetric porousmembrane. Optionally, the separation material is a mesh. Optionally, theseparation material comprises polyethylene (coated by ethylene vinylalcohol copolymer). Optionally, at least a portion of the separationmaterial comprises a polyethersulfone. Optionally, at least a portion ofthe separation material comprises an asymmetric polyethersulfone.Optionally, at least a portion of the separation material comprisespolyarylethersulfone. Optionally, at least a portion of the separationmaterial comprises an asymmetric polyarylethersulfone. Optionally, atleast a portion of the separation material comprises a polysulfone.Optionally, the separation material comprises an asymmetric polysulfone.Optionally, the separation material comprises a cellulose or cellulosederivative material. Optionally, the separation material comprisespolypropylene (PP). Optionally, the separation material comprisespolymethylmethacrylate (PMMA).

In one non-limiting example, a bodily fluid separation material isprovided comprising a formed component capture region having ananti-hemolytic surface layer; a bodily fluid pass-through regioncomprising pass-through openings sized so that formed components do notenter the bodily fluid pass-through region and a reduced amount of fluidleaching material relative to than the capture region, wherein duringseparation material use, bodily fluid enters the capture region prior toentering the pass-through region.

In one non-limiting example, a bodily fluid separation material isprovided comprising an anti-hemolytic, formed component capture region;a bodily fluid pass-through region comprising pass-through openingssized so that formed components do not enter the bodily fluidpass-through region and having a reduced amount of anti-hemolyticmaterial relative to the capture region, wherein during separationmaterial use, bodily fluid enters the capture region prior to enteringthe pass-through region.

In one non-limiting example, a bodily fluid separation material isprovided comprising a first filter region of the separation materialhaving an anti-hemolytic coating and pore spacing sized to constrainformed blood components therein; a second filter region of theseparation material having pore spacing smaller than pore spacing of thefirst filter region with pores sized so that formed components do notenter the second filter region and configured to have an amount ofanti-hemolytic coating less than that of the first region.

In one non-limiting example, a bodily fluid separation material isprovided comprising a percolating network configured to capture formedblood components: a first region of the percolating network with ananti-hemolytic coating on structures in the region, said network withopenings sized and spaced to allow formed blood components to enter thefirst region but constraining blood components therein from passingcompletely through the first region; a second region of the percolatingnetwork with a reduced anti-hemolytic coating on structures sized andspaced to prevent formed blood components from entering the secondregion; wherein bodily fluid passes through the first region prior toreaching the second region.

One or more of the embodiments described herein may include one or moreof the following features. By way of non-limiting example, a separationmaterial may be a mesh. Optionally, at least a portion of the separationmaterial comprises a polyethersulfone. Optionally, at least a portion ofthe separation material comprises an asymmetric polyethersulfone.Optionally, at least a portion of the separation material comprisespolyarylethersulfone. Optionally, at least a portion of the separationmaterial comprises an asymmetric polyarylethersulfone. Optionally, atleast a portion of the separation material comprises a polysulfone.Optionally, the separation material comprises an asymmetric polysulfone.Optionally, the anti-hemolytic material on the separation materialcomprises single and/or double alkyl chain N-oxides of tertiary amines(NTA). Optionally, the first region comprises a first separationmaterial layer and the second region comprises a second separationmaterial layer. Optionally, the separation material comprises a firstseparation material coupled to a second separation material. Optionally,the separation material comprises at least two separate separationmaterials. Optionally, at least another region of the separationmaterial between the first region and the second region. Optionally, thefirst region is in fluid communication with the second region.Optionally, the first region is spaced apart from the second region.

In one non-limiting example, a method of forming a bodily fluidseparation material is provided comprising coating the separationmaterial with an anti-hemolytic coating on a first region and a secondregion of the separation material; reducing anti-hemolytic effect of thesecond region of the separation material relative to the first region,wherein when the separation material is in operation, bodily fluidpasses through the first region prior to reaching the second region.

In one non-limiting example, a method of forming a bodily fluidseparation material is provided comprising coating at least a firstregion of the separation material with an anti-hemolytic coating; notcoating at least second region of the separation material with theanti-hemolytic coating.

One or more of the embodiments described herein may include one or moreof the following features. By way of non-limiting example, reducing theanti-hemolytic effect may comprise washing off at least a portion of theanti-hemolytic coating on the second region. Optionally, washing offcomprises directing solvent through the separation material. Optionally,washing off comprises soaking only a portion of the separation materialin a solvent. Optionally, the anti-hemolytic effect comprises addinganother coating of a different material over the anti-hemolytic coatingon the second region. Optionally, reducing the anti-hemolytic effectcomprises treating the separation material to bring its electricalcharge state to a neutral state and thus reduce the attraction of ionsthat increase the anti-hemolytic effect.

Optionally, the device has a distributor positioned with the separationmaterial to define an interface provides for propagating the samplelaterally over the separation material relative to a long longitudinalaxis of separation. Optionally, the device further comprises a plenumcoupled to the sample inlet, wherein the distributor is coupled atmultiple locations to the plenum, and wherein the plenum has a plenumoutlet for outputting sample not passing through the separation materialsuch that the device can output separated sample from the at least oneoutlet that has passed through the separation material and output samplefrom the plenum outlet that has not passed through the separationmaterial

In one non-limiting example, a device is provided for collecting asample from a subject and outputting a filtrate from the sample.

In one non-limiting example, a device is provided device for collectinga sample from a subject and forming a filtrate from at least a portionof the sample.

In one non-limiting example, a method is provided of using a device forcollecting a sample from a subject and outputting a filtrate from thesample.

In one non-limiting example, a method is provided of processing a formedcomponent separation membrane.

In one non-limiting example, a method is provided of processing a formedcomponent separation material.

In one non-limiting example, a method is provided comprising using aseparation material coupled to a housing to separate a formed componentportion of the sample from a liquid portion of the sample.

Optionally, a method is provided comprising at least one technicalfeature from any of the prior disclosed features. Optionally, a methodis provided comprising at least any two technical features from any ofthe prior disclosed features. Optionally, a device comprising at leastone technical feature from any of the prior disclosed features.Optionally, device comprising at least any two technical features fromany of the prior disclosed features. Optionally, a system comprising atleast one technical feature from any of the prior disclosed features.Optionally, a system comprising at least any two technical features fromany of the prior disclosed features.

This Summary is provided to introduce a selection of concepts in asimplified form that are further described below in the DetailedDescription. This Summary is not intended to identify key features oressential features of the claimed subject matter, nor is it intended tobe used to limit the scope of the claimed subject matter.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.However, in the event of a conflict between the content of the presentexpress disclosure and the content of a document incorporated byreference herein, the content of the present express disclosurecontrols.

COPYRIGHT

This document contains material subject to copyright protection. Thecopyright owner (Applicant herein) has no objection to facsimilereproduction of the patent documents and disclosures, as they appear inthe US Patent and Trademark Office patent file or records, but otherwisereserves all copyright rights whatsoever. The following notice shallapply: Copyright 2013-16 Theranos, Inc.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a schematic view of a device according to one embodimentdescribed herein.

FIGS. 2 to 4 show various views of a device according to one embodimentdescribed herein.

FIGS. 5A-5B show top-down plan views of devices according to embodimentsdescribed herein.

FIGS. 6-7 show various views of a device according to one embodimentdescribed herein.

FIGS. 8-9 show various views of a device according to one embodimentdescribed herein.

FIGS. 10-11 show various views of a device having at least two samplepathways according to one embodiment described herein.

FIGS. 12-13 show various views of a device according to one embodimentdescribed herein.

FIGS. 14-19 show cross-sectional views of various configurations forsample inlet openings and channels according to embodiments herein.

FIGS. 20-21 show views of various configurations for sample inletsaccording to embodiments herein.

FIGS. 22-28 show various patterns for sample distribution pathwaysaccording to embodiments herein.

FIGS. 29-30 show various views of a device according to one embodimentdescribed herein.

FIGS. 31-32 show various views of a device according to one embodimentdescribed herein.

FIGS. 33-34 show various views of a device according to one embodimentdescribed herein.

FIG. 35 shows various views of geometric configuration for the separatoraccording to embodiments described herein.

FIGS. 36-38 show one non-limiting example of sample inlet flow over theseparator according to embodiments described herein.

FIGS. 39-42 show one non-limiting example of sample outlet flow from theseparator according to embodiments described herein.

FIGS. 43-44 show side cross-sectional views of embodiments describedherein.

FIGS. 45-46 show top down plan views of vents according to embodimentsdescribed herein.

FIGS. 47A-48B show various views of sample wetting of a separatoraccording to embodiments described herein.

FIGS. 49-51 show top down plan views of various distribution channelpatterns over the separator according to embodiments described herein.

FIG. 52 shows cross-sectional views of various channel patterns andshapes over and under the separator according to embodiments describedherein.

FIGS. 53-55 show non-limiting examples of various aspect ratios of theseparator according to embodiments described herein.

FIG. 56 shows a side cross-sectional view of one non-limiting example ofan exit pathway according to embodiments described herein.

FIGS. 57 to 59 show views of non-limiting examples of devices having atleast two sample pathways according to embodiments described herein.

FIG. 60 shows yet another configuration of a device according toembodiments herein.

FIG. 61 shows one non-limiting example of a cartridge having a samplecollector and sample separator according to embodiment herein.

FIGS. 62 to 77B show still further embodiments as described herein.

FIG. 78 shows a side, cross-sectional view of a separation materialaccording to one embodiment described herein.

FIG. 79 shows an exploded side, cross-sectional view of separationmaterials according to one embodiment described herein.

FIG. 80 is schematic of a multi-layer separation material according toone embodiment described herein.

FIGS. 81 and 82 illustrate methods according to embodiments describedherein.

FIG. 83 shows one mode of operation according to at least one embodimentdescribed herein.

FIG. 84 shows a cross-sectional view of one embodiment of a device shownin FIG. 27D.

FIGS. 85A to 85D show various sample propagation patterns.

FIGS. 86 to 94 show still further embodiments of devices as describedherein.

FIGS. 95A to 95D show various embodiments of channel layouts for adistributor as described herein.

FIGS. 96 to 97 show various embodiments of channel layouts for adistributor as described herein.

FIG. 98 shows a perspective cross-sectional view showing a “lower”portion of one embodiment of a device described herein.

FIG. 99 shows an enlarged close-up view of one cross-sectional viewshowing a compressed membrane portion and structures beneath themembrane as described herein.

DESCRIPTION OF THE SPECIFIC EMBODIMENTS

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory onlyand are not restrictive of the invention, as claimed. It may be notedthat, as used in the specification and the appended claims, the singularforms “a”, “an” and “the” include plural referents unless the contextclearly dictates otherwise. Thus, for example, reference to “a material”may include mixtures of materials, reference to “a compound” may includemultiple compounds, and the like. References cited herein are herebyincorporated by reference in their entirety, except to the extent thatthey conflict with teachings explicitly set forth in this specification.

In this specification and in the claims which follow, reference will bemade to a number of terms which shall be defined to have the followingmeanings:

“Optional” or “optionally” means that the subsequently describedcircumstance may or may not occur, so that the description includesinstances where the circumstance occurs and instances where it does not.For example, if a device optionally contains a feature for a samplecollection unit, this means that the sample collection unit may or maynot be present, and, thus, the description includes both structureswherein a device possesses the sample collection unit and structureswherein sample collection unit is not present.

As used herein, the terms “substantial” means more than a minimal orinsignificant amount; and “substantially” means more than a minimally orinsignificantly. Thus, for example, the phrase “substantiallydifferent”, as used herein, denotes a sufficiently high degree ofdifference between two numeric values such that one of skill in the artwould consider the difference between the two values to be ofstatistical significance within the context of the characteristicmeasured by said values. Thus, the difference between two values thatare substantially different from each other is typically greater thanabout 10%, and may be greater than about 20%, preferably greater thanabout 30%, preferably greater than about 40%, preferably greater thanabout 50% as a function of the reference value or comparator value.

As used herein, a “sample” may be but is not limited to a blood sample,or a portion of a blood sample, may be of any suitable size or volume,and is preferably of small size or volume. In some embodiments of theassays and methods disclosed herein, measurements may be made using asmall volume blood sample, or no more than a small volume portion of ablood sample, where a small volume comprises no more than about 5 mL; orcomprises no more than about 3 mL; or comprises no more than about 2 mL;or comprises no more than about 1 mL; or comprises no more than about500 μL; or comprises no more than about 250 μL; or comprises no morethan about 100 μL; or comprises no more than about 75 μL; or comprisesno more than about 50 μL; or comprises no more than about 35 μL; orcomprises no more than about 25 μL; or comprises no more than about 20μL; or comprises no more than about 15 μL; or comprises no more thanabout 10 μL; or comprises no more than about 8 μL; or comprises no morethan about 6 μL; or comprises no more than about 5 μL; or comprises nomore than about 4 μL; or comprises no more than about 3 μL; or comprisesno more than about 2 μL; or comprises no more than about 1 μL; orcomprises no more than about 0.8 μL; or comprises no more than about 0.5μL; or comprises no more than about 0.3 μL; or comprises no more thanabout 0.2 μL; or comprises no more than about 0.1 μL; or comprises nomore than about 0.05 μL; or comprises no more than about 0.01 μL.

As used herein, the term “point of service location” may includelocations where a subject may receive a service (e.g. testing,monitoring, treatment, diagnosis, guidance, sample collection, IDverification, medical services, non-medical services, etc.), and mayinclude, without limitation, a subject's home, a subject's business, thelocation of a healthcare provider (e.g., doctor), hospitals, emergencyrooms, operating rooms, clinics, health care professionals' offices,laboratories, retailers [e.g. pharmacies (e.g., retail pharmacy,clinical pharmacy, hospital pharmacy), drugstores, supermarkets,grocers, etc.], transportation vehicles (e.g. car, boat, truck, bus,airplane, motorcycle, ambulance, mobile unit, fire engine/truck,emergency vehicle, law enforcement vehicle, police car, or other vehicleconfigured to transport a subject from one point to another, etc.),traveling medical care units, mobile units, schools, day-care centers,security screening locations, combat locations, health assisted livingresidences, government offices, office buildings, tents, bodily fluidsample acquisition sites (e.g. blood collection centers), sites at ornear an entrance to a location that a subject may wish to access, siteson or near a device that a subject may wish to access (e.g., thelocation of a computer if the subject wishes to access the computer), alocation where a sample processing device receives a sample, or anyother point of service location described elsewhere herein.

As used herein, the term “separator” may include a mesh, a filter, amembrane, a porous membrane, an asymmetric porous membrane, asemipermeable hollow fiber membrane, a percolating network structure, amaterial that can be used for size-exclusion of objects greater than acertain dimension, or other filtering material. Materials useful for thepreparation of the separating material may be selected from the groupcomprising polyethylene (coated by ethylene vinyl alcohol copolymer),polyacrylates, polystyrene, polyethylene oxide, cellulose, cellulosederivatives, polyethersulfone (PES), polypropylene (PP), polysulfone(PSU), polymethylmethacylate (PMMA), polycarbonate (PC),polyacrylonitrile (PAN), polyamide (PA), polytetrafluorethylene (PTFE),cellulose acetate (CA), regenerated cellulose, and blends or copolymersof the foregoing, or blends or copolymers with hydrophilizing polymers,including with polyvinylpyrollidone (PVP) or polyethyleneoxide (PEO).Suppliers of such materials and/or membranes include but are not limitedto BASF, Advanced Microdevices P. Ltd., International Point of CareInc., Gambro Lundia AB, Asahi Kasei Kuraray Medical Co., Ltd., GEHealthcare (Whatman division), or the like.

As used herein, the terms “sample” and “biological sample” refer to ablood, urine, sputum, tears, material(s) from a nasal swab, throat swab,cheek swab, or other bodily fluid, excretion, secretion, or tissueobtained from a subject. These terms are inclusive of an entire sampleand of a portion of a sample. As used herein, reference to a fluidsample includes reference to a sample and a biological sample. Suchsamples may include fluids into which material has been deposited, wheresuch material may be obtained from a nasal swab, throat swab, cheekswab, or other sample which may include solid or semi-solid material,whether along with or without natural fluids. Such fluids and samplescomprise fluid samples and sample solutions.

As used herein, the term “formed component” may include solid,semi-solid, or cellular structures such as but not limited to red bloodcells, white blood cells, platelet, or other components that may befound in a sample, a biological sample, bodily fluid, or natural fluid.

As used herein, the terms “fill” and “filled” and their grammaticalequivalents, e.g., as used in phrases such as “a vessel may be filledwith a sample solution” refer to the transfer of any amount, includingpartial filling and complete filling. These terms as used herein do notrequire that such filling completely fill a container, but include anylesser amount of filling as well.

It should be understood that the devices herein can be configured foruse with sample applied to the device, sample drawn into the device bycapillary force, sample delivered into the device by way of venipunture,sample delivered into the device by way of arterial puncture, nasalswab, tear collection, collection from any open wound, biopsy, or othersample delivery or acquisition technique and is not limited to anyspecific example described herein.

Referring now to FIG. 1, one embodiment of a formed component separationdevice will now be described. FIG. 1 shows a side cross-sectional viewof a device 10 having a formed component separator 20 positioned along apathway as indicated by arrow 30. In this non-limiting example, thedevice 10 has at least one sample inlet 40 for receiving a liquid samplethat has the formed components there and at least a first outlet 50 foroutputting only a liquid portion of the formed component liquid sample.As seen in FIG. 1, the pathway 30 fluidically couples the sample inletwith the first outlet. FIG. 1 also shows that sample such as but notlimited to blood flows from the inlet 40 and into and/or over the formedcomponent separator 20. In this non-limiting example, blood enters theformed component separator 20, where blood cells are trapped based onthe principle of size exclusion. The formed component separator 20, inone embodiment, may have a plurality of pores wherein those on onesurface are significantly smaller than those on another surface of theseparator 20. In this manner, the cells in the blood can enter theseparator 20 through the larger pores but cannot pass completely throughthe separator due to the much smaller pores on the output side of theseparator 20.

As the sample flows across the separator 20, the liquid portion of thesample such as but not limited to plasma is pulled away from the back ofthe separator 20 via a combination of capillary action and/or appliedpressure differential. Plasma flows away from the separator 20 asindicated by arrow 30. In one embodiment, the walls within the device 10may be coated with material such as but not limited to anti-coagulantfor mixing with the sample during filling.

Referring now to FIG. 2, another embodiment of a formed componentseparation device will now be described. In this non-limiting example,the device 100 has an inlet 102 that is open towards a top surface ofthe device 100. The inlet 102 is connected by a channel 104 to adistributor 110 that preferentially spreads the sample over theseparator 20. The liquid portion of the sample is outputted to thecollector 120 which may be directed to an external channel 130 such as aneedle or adapter channel. It should be understood that the distributor110 is not restricted to any particular structure or material and may bea plurality of capillary channels or tubes that distribute the sampleover the membrane. In some embodiments, it may be a hydrophilic coatingthat may be a continuous coating or a patterned coating to draw sampleto flow over the membrane.

Referring now to FIG. 3, a cross-sectional view of one portion of thedevice 100 (as indicated by arrows 3-3 in FIG. 2) shows some of thedetails regarding this embodiment of the distributor 110 thatpreferentially spreads the sample over the separator 20, and thecollector 120 which draws liquid away from the separator 20. Sample willflow across the separator 20 as indicated by arrow 122. In thisnon-limiting example, the lead-in channel 130 wicks blood in from theinlet 102 and transports it into the distribution channel network ofdistributor 110 via capillary action. The distribution channel networkcomprises a network of capillaries on the blood side of the separator20. This distributor 110 pulls sample away from the lead-in channel anddistributes it evenly over the membrane. In one embodiment, theseparator 20 separates plasma from whole blood via a two-step process.One process uses a passive mechanism: gravity and capillary forcegradient. A second process uses an active mechanism: application of apressure differential. These processes can act in a sequential manner orin a simultaneous manner. Capillaries of collector 120 also route theliquid portion of the sample into the extraction port when the pressuredifferential is applied. Sample flow through the device 100 is indicatedby arrow 140.

Referring now to FIGS. 5A and 5B, various embodiments of collectiondevices will now be described. FIG. 5A shows a sample separation device150 that has an aspect ratio that provides for fewer numbers of channelsbut increases the length of each of the channels.

The length of the membrane along a longitudinal axis of the device,relative to the width may be in a range of about 3:1 to about 5:1. FIG.5B shows another embodiment of a separation device 160 that has adifferent aspect ratio which an increased number of capillary channels,but reduced length for each. The length of the membrane along alongitudinal axis of the device, relative to the width may be in a rangeof about 1:1 to about 1:3. It should also be understood that thecross-sectional size of channels over separator 20 and those beneath theseparator 20 can also be different. In one embodiment, the channels inthe collector 120 that are beneath the separator 20 are at least 2×smaller in cross-sectional area than those over the channel. In oneembodiment, the channels in the collector 120 that are beneath theseparator 20 are at least 5× smaller in cross-sectional area than thoseover the channel. In one embodiment, the channels in the collector 120that are beneath the separator 20 are at least 10× smaller incross-sectional area than those over the channel. The decreased size ofthe channels will increase the capillary pressure and thuspreferentially direct liquid portions of the sample towards the outputof the device.

Referring now to FIGS. 6 and 7, yet another embodiment of a sampleseparation device 170 is shown. FIG. 6 shows a top-down view of a bottomportion of the separation device 170 is shown with a vent 172, a ventinlet channel 173, and a collector 176 is shown with a plurality ofchannels to draw sample from an underside of the separator 20 (moreclearly shown in FIG. 7). An outlet tube 178 such as but not limited toa needle can be used to engage a container such as but not limited to asealed container with piercable septum or cap, wherein the interior orthe container is under vacuum pressure therein to pull liquid sampleinto the container when it is fluidically engaged by the needle of theoutlet tube 178. Optionally, the container may take the form of a testtube-like device in the nature of those marketed under the trademark“Vacutainer” by Becton-Dickinson Company of East Rutherford, N.J.

FIG. 7 shows, in one embodiment, a side cross-sectional view of thedevice 170. As can be seen, the separator 20 is “sandwiched” between thedistributor 174 and the collector 176. The separation material along thefirst pathway configured to remove formed components from the sampleprior to outputting at the first outlet. Processed sample will beoutputted through the outlet tube 178 into a container or otherreceptacle. Some embodiments as seen here may have a funneled portion inthe collector 176 to direct sample that has been processed towards theoutlet tube 178. By way of example and not limitation, the sample can beapplied directly to the distributor 174 or directly to the separator 20.

Referring now to FIGS. 8 and 9, a still further embodiment will now bedescribed. FIG. 8 is a perspective view of a sample separation device190. This embodiment of the sample separation device 190 is configuredto allow for direct application of the sample onto the separator by wayof opening 192 over the separator. Processed liquid will be drawn by thecollector 194 to be output through the outlet 196.

Referring now to FIG. 10, a sample collection device 200 according toone embodiment herein will now be described. In this non-limitingexample, the sample collection device 200 includes a first pathway 202that is configured to direct sample to a separator 204. The samplecollection device 200 also includes a second pathway 206 that collectssample but does not direct it through a formed component separator 204.Both pathways 202 and 206 have openings that are co-located, adjacent,coaxial, or otherwise closely positioned at a distal end 208 of thedevice 200 that will be in contact with the subject. Optionally, someembodiments may share a common pathway that has a single opening at thedistal end 208. The collected sample may exit from one or more adapterchannels 210 to one or more sample containers (not shown for ease ofillustration). FIG. 10 shows that the distributor 212 may have featuresthat extend beyond the area of the separator 204. These off-membranefeatures are helpful in drawing the sample towards and over themembrane, particularly as the channel widens to accommodate themembrane.

FIG. 11 is a cross-sectional view of one embodiment of the distributor212 over the separator 204 as indicated by arrows 11-11 in FIG. 10. Asseen in FIG. 11, at least a portion of the separator 204 may havereduced thickness area 214 where the material may be thinner oroptionally where the material is compressed from its normal thickness tohold the material in place. In one non-limiting example, one purpose ofthis compressed region is to compress the pores in the membrane andthereby create a seal that is impassable by the formed components. Inone non-limiting example, normal separator thickness may be in range ofabout 100 to about 1000 microns. Optionally, normal separator thicknessmay be in range of about 200 to about 900 microns. Optionally, normalseparator thickness may be in range of about 200 to about 500 microns.Optionally, normal separator thickness may be in range of about 300 toabout 500 microns. Optionally, normal separator thickness may be inrange of about 300 to about 800 microns. Optionally, normal separatorthickness may be in range of about 400 to about 700 microns. Optionally,normal separator thickness may be in range of about 500 to about 600microns. FIG. 11 also shows that the collector 216 may be a plurality ofcapillary channels that have a v-shaped cross-section. These are used todraw the liquid only sample to the outputs of the device at adapterchannel 210.

Referring now to FIGS. 12 and 13, a still further embodiment of a samplecollection and separator device 220 will now be described. FIG. 12 showsthe device 220 as having an inlet 222 for receiving sample as indicatedby arrow 224. The sample received at inlet 222 enters a channel 226 thatis aligned along an axis configured to intersect the plane in which theseparator 228 is positioned. In this manner, the sample when it contactsthe separator 228 is placed onto primarily a planar surface of theseparator 228. In one embodiment, the peripheral portion 230 ofseparator 228 is compressed to hold the separator in place and toprevent sample from exiting along the edge of the membrane, instead ofthrough the back and into the collector 232.

At the point of sample contact with the separator 228 and the end ofchannel 226, the sample contact both the separator 228 and channels 234of the distributor 236. In this manner as will be discussed in moredetail elsewhere herein, the sample is drawn by both the separator andthe distributor 236 to be distributed over and/or through the separator.Optionally, this can be beneficial to prevent clogging of sample at anyone location or junction point on the separator. Optionally, thedistributor may be used to facilitate longitudinal uniformity of thesample with respect to concentration of formed components in the liquidportion of the sample. Optionally, use of the distributor 236 can alsospeed the filling process. The channels 234 may be coupled to one ormore vents 238 that allow for gas or air to be displaced when sampleenters the distributor 236. FIG. 12 shows that each channel 234 may haveits own individual vent 238. Optionally, some embodiment may have two ormore channels 234 couple to share a vent by way of common manifoldconfiguration or the like. As seen, the vents are positioned at the endsof the channels 234 to allow for the channels to fully fill.

FIG. 13 shows a lateral cross-sectional view of one embodiment thedevice 220 wherein the channels 234 of the distributor 236 are shownover the capillary collection channels 240 of the collector 242. FIG. 13also shows that not ever channel in the collector 242 has the samecross-sectional shape. By way of non-limiting example, the channels 244along a perimeter of the collector 242 may have a different shape suchas but not limited to a rectangular cross-section that is different fromother channels in the collector 242.

Referring now to FIG. 14, a cross-sectional view of one embodiment of asample inlet channel will now be described. As seen in FIG. 14, theinlet channel 226 directs sample from inlet 222 towards the separator232. The angled orientation of channel 226 relative to the plane of theseparator 232 allows for sample to be placed onto the planar surface ofthe separator and not relying purely on lateral pulling. The angledcross-sectional shape also increases the area of sample contact to begreater than merely the lateral cross-section of the channel.

FIGS. 15 and 16 also show other embodiments wherein sample inletchannels 250 and 252 that have sample channel transition features 254and 256 that minimize detrimental effects due to change in channeldimension. These transitions features may be configured to reducedimension in one axis (feature 254) or minimize a sudden change indimension (feature 256) by gradually transitioning the change indimension over a longer and/or wider area. It should be understood thatsome embodiments may combine the use of features 254 and 256. Otherembodiments herein may also have these features or others that useembody the concepts described herein to minimize detrimental impact ofcertain channel features. Inlet channel desirably leads to directcontact with the membrane, which in one non-limiting example, is withoutan intermediary reduction in capillary forces, which can stop the bloodflow and prevent distribution.

Referring now to FIGS. 17 to 19, other configurations for sample inletchannels according to embodiments herein will now be described. FIG. 17shows an angled sample inlet channel 260 that has a “splitter”configuration wherein at least one opening of channel 262 couples withthe inlet channel 260 to direct a portion of the sample to channel 262.This can be particularly useful in configurations such as but notlimited to that shown in FIG. 10 wherein one portion of the sample willbe treated to separate formed components from the liquid portion of thesample while other portions of the sample are not treated in the samemanner and thus progress down one or more other pathways.

In this non-limiting example, the opening for channel 260 is at least aslarge as if not larger than the cross-sectional shape of the inletchannel 260. The sample continues in a second portion 264 of the inletchannel 260 to reach the separator 232. The openings 266 of adistributor for the separator 232 can be located at the end portion ofthe channel 260. As seen in this non-limiting example, the secondportion 264 of channel 260 is smaller in cross-sectional area than aninitial portion of the channel 260. FIG. 17 also shows that the channel260 is directing sample at an upper portion of the channel profile tothe second portion 264 while the opening for channel 262 collect atleast sample in the lower portion of the channel profile. A higher entrypoint can help with lengthwise blood distribution along the length ofthe separator by delaying and reducing initial penetration of theseparator by the sample as it flows into the distribution volume.

FIG. 18 shows yet another embodiment of the sample inlet channel whereinthe opening for channel 263 is now configured to interface only asmaller portion of the channel 260. As seen in FIG. 18, the opening 263of the channel intersects only a lower portion of the channel profilefor channel 260. This can useful to customize the volume of sample thatis directed towards each channel.

FIG. 19 shows yet another embodiment wherein the inlet channel 270connects to the second channel 272 and has a significantly largercross-sectional profile relative to the second portion 274 of the inletchannel. The second portion 274 is configured to draw sample from anupper portion of the cross-sectional profile of the inlet channel. Theopenings 276 for the sample distributor draw from the lower portion ofthe portion 274 to distribute sample over the separator 232. A collector278 will draw liquid sample from the separator 232. At least one vent280 can be coupled to the separator 232 to provide a controlled inlet ofexternal atmosphere to facilitate the pull of liquid device. In onenon-limiting example, vent 280 may allow at least some venting to occurduring the dynamic stage of extraction, in which a pressure differentialor other motive force is applied to draw the liquid portion of thesample into at least one collection container. Optionally, the vent 280may be separated from the collector 278 by the separator 232 to providea controlled inlet. The vent 280 may couple to compressed portion of theseparator 232. The vent 280 may couple to normal portion of theseparator 232.

Referring now to FIGS. 20 and 21, various shapes can be configured foruse to engage a subject for sample collection according to embodimentsherein. FIGS. 20 and 21 both show protrusions for use on collectiondevices as described herein. FIG. 20 shows a protrusion 290 that isshaped in a scoop or spoon configuration having both vertical andhorizontal portions of the opening 292 in the protrusion accessible tothe user to collect sample. The opening 292 may lead to a single ormultiple pathways in the device.

FIG. 21 shows one embodiment of a protrusion 294 that extends away fromthe body of the device so that the user is provided a visual cue as towhere the contact the device to the subject to collect sample. Theopening may be funnel shaped to assist in sample collection and inengagement with the skin of the patient. The protrusion 294 has anopening that may lead to a single or multiple pathways in the device. Itshould be understood that some embodiments may have protrusions that areshaped to be convex or concave to facilitate engagement of the deviceprotrusion with bead or droplet of bodily fluid sample on the subject.The protrusion may be coated with hydrophilic and/or hydrophobicmaterial to push or pull the sample in a desired direction.

Referring now to FIGS. 22 to 24, it should also be understood thatsample can be delivered to one or more different locations on the sampleseparator according to at least one embodiment herein. As seen in FIGS.19 to 21, some embodiments may deliver sample to one end of theseparator, away from a central portion of the separator. Optionally,some embodiments as seen in FIGS. 22 to 23, deliver sample from an inletat one end to one or more openings closer to the center of theseparator. The sample may be delivered both at the one end and atlocations closer to the center.

For example, FIG. 22 shows embodiments of inlet tubes 310, 312, and 314for use in delivering sample from an inlet on a periphery of the deviceto one or more locations along a central area of the separator 316. Thelocations 320, 322, and 324 may be openings or other structures thatallow the inlet tubes 310, 312, and 314 to deliver sample to the desiredlocation on the separator 316. FIG. 22 shows that these locations may bedistributed over various locations on the separator 316. FIG. 23 showsan embodiment wherein the locations 330, 332, and 334 are located in aline near the central area of the separator. Optionally, someembodiments may use single or multiple combinations of one or more ofthe structures in FIGS. 19 to 24 to provide a desired sampledistribution pattern over the separator. It should be understood thatthe embodiments herein can deliver the sample directly onto theseparator, onto network of distributor channels over the separator, or acombination of the foregoing.

FIG. 24 shows a still further embodiment wherein the inlet is notlocated at either end of the collection device, but instead has an inletthat is substantially centrally located as seen in FIG. 24. Thisembodiment shows that the inlet 340 leads to a channel 342 that feeds toa central portion of the separator 344. FIG. 24 is a simplified drawingshowing primarily only the separator 344 and an outlet port 346 thatdraws liquid portion of the sample away from the separator afterprocessing.

Referring now to the embodiments of FIGS. 25 and 26, it should beunderstood that the distribution of the channels of the distributor isnot limited to the patterns, sizes, or shapes disclosed in the previousfigures. As seen in FIG. 25, one embodiment may align all of thechannels 350 orthogonal to the longitudinal axis of the device and/orseparator. In the embodiment of FIG. 25, this results in a greaternumber of channels 350, but each has a shorter length. Optionally, theorientation of the channels is not limited to orthogonal to thelongitudinal axis of the device. Other angles relative to thelongitudinal axis of the device and/or the separator are not excluded.Optionally, some embodiments may use different patterns over differentportions of the separator. Optionally, some embodiments can use acombination of patterns over the same area.

It should also be understood that this same or similar pattern ofchannels can also be implemented on the collector that is used on theopposite side of separator. Optionally, the distributor can use onechannel pattern and the collector can use a different channel pattern.

Optionally, some of the sideways capillaries 350 on the collector uses anon-vented configuration. Some of these sideways capillaries 350demonstrated a different extraction behavior as blood separates.Lengthwise capillaries tend to extract from back of device first, thentowards the front. Sideways capillaries 350 extract first towards themiddle of the separator and outwards towards front and back of device,which can be used to create a more even extraction process across theseparator.

As seen in FIG. 26, some embodiments may also use a configuration havinga manifold 360 having a plurality of outlets 362 that can distributesample over the separator and/or into the distributor. Some embodimentscan have shorter or longer outlets 362, depending on the pattern thatone desires to deliver sample to the separator and/or distributor. Itshould also be understood that some embodiments may more than onemanifold 360 that delivers sample to the separator and/or distributor.For example, one embodiment may have another manifold 360 deliver samplealong the other longitudinal edge of the separator.

FIG. 26 also shows in phantom a potential pattern for a collectionmanifold 364 for use on the opposite side of the separator for samplecollection. This manifold 364 would typically not be used on the sameside as the distribution manifold 360 but would instead be on anopposite side of the separator.

Referring now to FIG. 27A, a still further embodiment is shown with apatterned manifold 370 with channels that distribute sample alongvarious locations over the separator 372. FIG. 27A shows that there maybe a plurality of vents 374 that allow for gas or air in the separator372 or other part of the manifold to escape as sample fills the area. Itshould also be understood that although the manifold 370 is shown with adistribution pattern of substantially similar length channels 376, suchchannels can be pattern to have same, different, repeating, or otherpatterns of size, length, contact area with the separator, or otherdimension to provide a desired performance. It should also be understoodthat the manifold 370 can be used to directly distribute sample onto theseparator or it may opt to deliver sample in a pattern to a distributorwhich then further distributes the sample over the separator. Someembodiments of the manifold 370 uses tubes with openings at selectlocations to allow sample to exit. Some embodiments can use channelswith at least one open side to distribute sample along a certain lengthof the separator and/or distributor.

Referring now to FIG. 27B, a still further embodiment is shown with apatterned manifold 371 with channels that distribute sample alongvarious locations over the separator 372. It should also be understoodthat although the manifold 371 is shown with a distribution pattern ofsubstantially channels 377, such channels can be pattern to have same,different, repeating, or other patterns of size, length, contact areawith the separator, or other dimension to provide a desired performance.Relative the embodiment of FIG. 27A, this embodiment with manifold 371uses shorter length channels 377 as compared to channels 376 of manifold370. FIG. 27B also shows that embodiments of the manifold 371 mayinclude multiple longer length channels 378 to distribute sample to theintersection channels 377. FIG. 27B shows there are three channels 378,but it should be understood that other embodiments may have a differentnumber of channels. It should also be understood that the manifold 371can be used to directly distribute sample onto the separator or it mayopt to deliver sample in a pattern to a distributor which then furtherdistributes the sample over the separator. Some embodiments of themanifold 371 uses tubes with openings at select locations to allowsample to exit. Some embodiments can use channels with at least one openside to distribute sample along a certain length of the separator and/ordistributor.

FIGS. 27C and 27D show a still further embodiment with a manifolddesigned to have an aspect ratio where the distribution pattern of thechannels is configured to pass along a short dimension of the separatorversus another longer dimension. It should be understood that this typeof distribution pattern may be modified for use in any of theembodiments described herein, such as but not limited to those of FIG.77A-77B.

Referring now to FIG. 28, yet another embodiment of a manifold 380 isshown. This can be as a distributor that has twelve channels thatdistribute sample over the separator. As seen, the channel pattern ofmanifold 380 initially has six channels leading away from a single inletchannel, and those six channels are each split once to achieve twelvechannels.

Referring now to FIGS. 29 to 34, still other embodiments showingdifferent combinations of inlet channels and distributors are shown.FIGS. 29 and 30 show a single inlet 390 having a circularcross-sectional shape leading to a multi-channel distributor 392 withchannels 394, with each of the channels coupled to its own vent 396,similar to that shown for FIG. 12. It should be understood, however,that embodiments where vents are shared are not excluded. FIG. 30 showsthe cross-sectional shape of the channels 394 and their size relative tothe capillary channels 398 of the liquid sample collector.

FIGS. 31 and 32 show at least one embodiment of an inlet channel 400having a low aspect ratio in terms of channel height to width. Thenarrow height, wide inlet channel 400 leads to a multi-channeldistributor 402, wherein the channels 404 also have low height to widthaspect ratios and also have intersecting connectors 406 that provideconnector pathways between the channels to form a grid or other pattern.In this particular embodiment, the connectors 406 are pathways withnarrower cross-sectional areas that of the channels 404. Each of thechannels 404 is coupled to its own vent 408, but it should be understoodthat embodiments where vents are shared are not excluded. The low aspectratio of the channels 404 are more clearly shown in FIG. 32 along withtheir cross-sectional area relative to the cross-sectional area of thechannels 409 of the collector.

FIGS. 33 and 34 show an embodiment having an inlet 420 comprising aplurality of individual channels 422 that are co-located as the inlet420. Once sample is collected, each of inlet channels 422 directs itsportion of the sample to the distributor 424, which in this case is amulti-channel distributor, wherein the channels 426 have intersectingconnectors 428 that provide connector pathways between the channels toform a grid or other pattern. In this particular embodiment, theconnectors 428 are pathways with at least the same or greatercross-sectional area than that of the channels 426. Each of the channels426 is coupled to its own vent 429, but it should be understood thatembodiments where vents are shared are not excluded. The low aspectratio of the channels 426 are more clearly shown in FIG. 34 along withtheir cross-sectional area relative to the cross-sectional area of thechannels 430 of the collector. As seen in FIGS. 29-34, the separatorsare shown with its upper surface in contact at location 427 with a wallsurface of the device so that there is no gap. Some embodiments may havethe separator under compression to maintain this contact and to accountfor any variation due manufacturing tolerances. This contact may also betrue for the surfaces below the separator. By way of non-limitingexample, this vertical compression of the separator to overcome anymanufacturing tolerances can be applied to any of the embodimentsdiscussed or suggested herein.

Referring now to FIG. 35, it should be understood that the separatorshown in the embodiments up to this point have been rectangular, racetrack, oval, or some combination of the foregoing. FIG. 35 shows thatother shapes are not excluded and that the separator may be materialshaped such as but not limited to elliptical, triangular, quadrilateral(e.g., square, rectangular, trapezoidal, parallelogram), pentagonal,hexagonal, heptagonal, octagonal, square, circular, star, other twodimensional patterns, or single or multiple combinations of theforegoing. It should also be understood that the separator may beconfigured to be in certain three dimensional configurations such as butnot limited to tubular, cylindrical, disc, pyramid, mesa, or the likecan also be adapted for use herein. By way of non-limiting example, someinlet locations 440 for sample distribution are shown for someembodiments. These are merely exemplary and other positioning of thenumber and location of inlets 440 are not excluded.

Sample Flow over Separator

Referring now to the non-limiting examples of FIGS. 36 to 38, it shouldbe understood that configuration wherein the channels 234 are open onone side to separator 232 allows for a multi-mode sample propagationpattern wherein at least a first portion is propagating laterally withinthe separator and a second portion is propagating through the channels234 of the distributor over the separator 232. In this non-limitingexample, the multi-mode sample propagation shows a leading edge 450 thatis ahead of the sample in the channels at filled surface 452, which canexhibit a meniscus type shape as seen in FIG. 36. The sample continuesto fill the separator 232 with the multi-mode sample propagation patternas seen in FIG. 37 until the fill is completed as seen in FIG. 38, whensample is filled in the channels to reach the vents 238. In someembodiments, the volume of sample in the channels is greater than thatin the separator 232 and this may account for part of the reason thatthe leading edge in the separator 232 may be moving ahead of that in thechannels 234.

Sample Collection from Separator

Referring now to the non-limiting examples of FIGS. 39 to 42, at leastone non-limiting example of sample flow during separation will now bedescribed. Although not shown in the illustrations, at the point whenthe device is in the minimum fill condition as seen in FIG. 39, thesample is ready to be engaged by a force to draw sample more completelythrough the separator 232. In non-limiting example, there has alreadybeen some liquid sample that has traversed though the thickness of theseparator 232 and has been pulled by capillary pressure from thecapillary channels 240 of collector 242 to fill at least some of thosechannels and “prime” the channels with liquid that can then be used aspart of the system to complete processing of the remaining sample heldin the channels of the distributor above the separator 232. As indicatedby arrow 460, a pulling force such as but not limited to full or partialvacuum in a sealed container like a vacutainer can be used to startmoving liquid only sample into the container. As long as there is no“meniscus” break or if such breaks are recoverable, the sample still inthe separator 232 or above it will begin to be drawn though the device.

As seen in FIG. 40, the pull of liquid in the direction of arrow 460 onthe underside of the separator 232 will also create a pull that drawssample laterally toward and/or downward into the separator 232. It isoften desirable that this flow be without destructive trauma to formedcomponents trapped in the separator 232, as the release of material frominside these formed components into the sample is generally undesirable.FIG. 40 shows that some sample that was in the inlet 222 has been drawninto the channels 234. Sample has begun to drain into the separator 232in the channels 234 closest to the edge near the pulling force indicatedby arrow 460. FIG. 41 also shows that the sample continues to be drawndownward and in the direction of arrow 460, there is also movement ofsample further away from the inlet 222. FIG. 42 shows that uponcompletion of the separation process, form components such as but notlimited to red blood cells that have been size-excluded from the sampleremain and leave a light red color on the separator 232.

Referring now to FIGS. 43 and 44, a side cross-sectional view is shownof various embodiments of a sample collection and sample separationdevice. FIG. 43 shows that maximum trans separator pressure occursclosest to the end of the device where the outflow 460 is occurring. Thefurther away from the area of outflow 460, the lesser the trans pressureacross the separator. This gradient can explain in part the flow patternseen in FIGS. 39-42.

As the separator beings to become clogged with formed components nearthe extraction end at arrow 460, flow has an increasing lengthwisecomponent. Lengthwise intra separator flow increases shear stress onRBCs, and this trauma leads to greater hemolysis, even at lowerpressures. Shorter, wider separator exhibit this effect in a manner thatis less pronounced, while the effect is more pronounced in separators ofgreater lengths.

Referring now to FIG. 44, one embodiment herein comprises at least oneor more vents 470 on the back side/collector side of the collector thatdecouples filtration from extraction. By providing a controlled inlet,the excessive force conditions that may cause excess shear stress itrelieved by the controlled inlet from a pathway different from thoseoccupied by the formed components, thus shifting pressure away fromthose components and still allowing for lateral liquid flow duringextraction. In one non-limiting example, the controlled venting isbalanced by having the pathway to reach the vent 470 pass through aportion of the separator 232. In one embodiment, this is a portion ofthe separator 232 is not filled with sample. Optionally, this is acompressed portion of the separator 232 not filled with sample. In thismanner, there will be some level of venting that creates a pathway forair to enter by way of the vent to relieve the pressure put on formedcomponents in the sample if there is no separate inlet. In oneembodiment, the resistance is substantially equal to the resistanceassociated with venting through the separator 232 filled with sample. Inone embodiment, the resistance is less than the resistance associatedwith venting through the separator 232 filled with sample. With the ventstructure, one can extract plasma with reduced risk of hemolysis whendealing with blood samples.

Referring now to non-limiting examples of FIGS. 45 and 46, top downviews of vent structures in the lower half of the device is shown. FIG.45 shows that in this embodiment, the vent 480 is coupled to a shapedpathway 482 that is configured to be in communication with the capillarychannels 240 of the collector 242. Some embodiment may include a valve,porous material, mesh material, reduced diameter pathway, or other flowreducing material to control the flow of air from the vent to theinterior of the collector 242. Some embodiments may also have the shapedpathway 482 be compressed with material from the separator (not shown)to slow the flow to the collector 242.

Referring now to FIG. 46, the vent 484 of this embodiment is coupled toa shaped pathway 486 that is configured to be in the area where theseparator material (shown in phantom by line 487) will cover it. Thecoverage may be in a compressed manner. Optionally, the coverage may bewithout substantial compression. The communication with the capillarychannels 350 of the collector are separated by a pre-selected distance488 from the shaped pathway 486 of the vent. In this manner, the pathwaytravels through at least that distance 488 of separator material (whichmay be gas porous) before air from the vent is able to be in fluidcommunication with the channels 350. This can be useful in someembodiments to regulate the rate in when venting occurs.

It should be understood that although the shaped pathways 482 and 486are shown as continuous pathways, they may optionally be a plurality ofdiscontinuous, discrete openings linked to a common vent or having theirown individual vents. In some embodiments, it is desirable to locate thevent near the end of the device distant from the end where liquid sampleis being pulled from the device. Some embodiments may combine one ormore components of FIGS. 45 and 46 together regarding venting andregulation of air through any such vent.

Referring now to non-limiting examples of FIGS. 47 and 48, these figuresshow cross-sectional views of the separator showing differentpercentages of saturation by the sample. As seen in FIG. 48A, thespacing of the channels of the distributor over the separator can beselected to increase separator saturation. FIG. 47A shows large channelsspaced farther apart yields lower saturation that a combination ofsmaller channels spaced closer together to achieve a more uniformsaturation pattern in the material.

As seen in the top down view in FIG. 47B, directed wetted area 490 ascompared to indirectly wetted area 492 can be configured to increaseoverall saturation of the separator. The directly wetted surface area inFIG. 47B relative to total surface area is about 30%. ChannelSA/V=DWA/V=5.0 for FIG. 47B.

FIG. 48B shows directed wetted area 492 as compared to indirectly wettedarea 496 can be configured to increase overall saturation of theseparator. The directly wetted surface area in FIG. 48B relative tototal surface area is about 60%. Channels in the new configuration havea larger ratio of directly wetted surface area (DWA) to volume V, andnearly twice the directly wetted area as a fraction of total surfacearea (SA), wherein V is the volume of the separator. This results in amore effective wetting of the membrane; takes in more liquid per surfacearea. Channel SA/V=DWA/V=6.3 for FIG. 48B. In one embodiment, thedesired range of channel surface area relative to the surface area ofthe separator on that side of the separator is in the range of about 35%to 70%. Optionally, the desired range of channel surface area relativeto the surface area of the separator on that side of the separator is inthe range of about 40% to 70%. Optionally, the desired range of channelsurface area relative to the surface area of the separator on that sideof the separator is in the range of about 50% to 60%. In one embodiment,the ratio of Channel SA/V which is also DWA/V is in the range of about 5to about 10. In one embodiment, the ratio of Channel SA/V which is alsoDWA/V is in the range of about 4.5 to about 9. In one embodiment, theratio of Channel SAN which is also DWA/V is in the range of about 5 to8. Optionally, the ratio of Channel SA/V which is also DWA/V is in therange of about 6 to 8. Optionally, the ratio of Channel SA/V which isalso DWA/V is in the range of about 5.5 to 7. Optionally, the ratio ofChannel SA/V which is also DWA/V is in the range of about 6 to 7.

Referring now to non-limiting examples of FIGS. 49 to 51, variouspatterns of channels for distribution over the separator are shown. FIG.49 shows an embodiment wherein there are no channels over the separator20. FIG. 50 shows an embodiment with ten channels. Although distributedsymmetrically about a longitudinal axis of the separator, it should beunderstood that other embodiments where channel size, distribution, orlength are not symmetrical about the longitudinal axis may be used. FIG.51 shows an embodiment with twenty two distribution channels.

FIG. 52 shows a plurality of cross-sections of the device showing thedistributor, separator, and collector. As seen, the channels 500 of thedistributor can be of various cross-sectional shapes such as but notlimited to elliptical, triangular, quadrilateral (e.g., square,rectangular, trapezoidal, parallelogram), pentagonal, hexagonal,heptagonal, octagonal, square, circular, star, other two dimensionalpatterns, oval, half-oval, half-circular, polygonal, or single ormultiple combinations of the foregoing. The lengthwise pathway shape canalso be configured such as to distribute sample in a desired manner overthe separator. The channels 510 of the collector can be of variouscross-sectional shapes such as but not limited to elliptical,triangular, quadrilateral (e.g., square, rectangular, trapezoidal,parallelogram), pentagonal, hexagonal, heptagonal, octagonal, square,circular, star, other two dimensional patterns, oval, half-oval,half-circular, polygonal, or single or multiple combinations of theforegoing. In one embodiment, the channels shapes of the distributor andthe collector may be the same or different. Some embodiment of thedistributor may have different shaped and/or sized channels in thedistributor to provide a certain desired sample distribution pattern.Some embodiment of the collector may have different shaped and/or sizedchannels in the collector to provide a certain desired sample collectionpattern.

FIGS. 53 to 55 show various non-limiting examples of different aspectratios for the separators for use with the device. FIGS. 53 and 55 alsoshow different aspect ratios for the distributor used with such adevice. In one embodiment, the separator has a configuration where theaspect ratios, defined as the length of the separator 520 (lengthwisealong the direction of flow, toward the extraction port as indicated byarrow 521) divided by its width along arrow 523 are in the range ofabout 1:1 to 3:1. Optionally, the aspect ratio may be in the range of1:1 to 5:1. Optionally, some embodiments may have aspect ratios in therange of 5:1 to 1:1. It should be understood that in these figures, thechannels 500 are shown over the separator 520. A common vent 530 whichis shown in FIGS. 53 and 54 can be also adapted for use with otherembodiments described herein. FIG. 55 shows a plurality of differentaspect ratios for the separator 520 and the distributor having channels500.

FIG. 56 shows one example of an exit conduit 540 below the collector 550that shows a round inner surface 542 in the 90 degree elbow thattransitions directions of sample flow out of the device from a verticalto a lateral flow.

FIGS. 57 to 59 show that in addition to the pathway 600 for separationof formed components from the sample, some embodiments of the device arealso configured to allow for other pathways 610, 620, or 630 thatcollect sample for treatment in a different manner. As seen in thefigures, these pathways can be shaped and sized so that they can containa desired amount of sample therein. Some embodiment may be configured sothat the pathlength is such that the fill times for both the formedcomponent separated sample and the un-separated sample are substantiallythe same. In this manner, a single indicator can be used to alert theuser that sufficient fill has been achieved in both pathways.

FIGS. 57 to 59 also show that the output of the devices may be intocontainers 660. In one non-limiting example, the container may be but isnot limited to a sealed container with piercable septum or cap, whereinthe interior or the container is under full, partial, or some level ofvacuum pressure therein to pull at least a certain volume of liquidsample into the container when it is fluidically engaged by the needleof the outlet tube or needle of the devices described herein.Optionally, the container may take the form of a test tube-like devicein the nature of those marketed under the trademark “Vacutainer” byBecton-Dickinson Company of East Rutherford, N.J. The output of onedevice may be both blood (B) and plasma (P). Optionally, the output canbe viewed as a) separated liquid-only sample and b) other sample output.Optionally, the output can be viewed as a) separated liquid-only sample(and any formed components smaller than the size exclusion limit) and b)other sample output. One or more of the pathways may be treated, coated,or otherwise prepared to deliver a material into the sample such as butnot limited to an anti-coagulant, ethylenediaminetetraacetic acid(EDTA), citrate, heparin, or the like as currently known or will bedeveloped in the future. Some may have two or more the pathways treatedwith the same or different material.

FIG. 59 shows a still further embodiment showing a Y-split to separatesample to go in to both pathways. It should be understood that althoughthis indication of fill level in one or more of the pathways may be byway of a visual indication. It should also be understood that otherindication methods such as but not limited to audio, vibratory, or otherindication methods may be used in place of or in combination with theindication method. The indicator may be on at least one of thecollection pathways. Optionally, indicators are on all of the collectionpathways. It should be understood that the devices herein can beconfigured to have three or more pathways and are not limited to onlytwo pathways.

For any of the embodiments herein, there can be container(s) such as butnot limited to container 660 for use in drawing liquid sample that hasgone through or will be drawn through the separator. In someembodiments, this is a two phase process, where there is an initialfilling phase of sample into the separator using a first motive forceand then a second phase using a second motive force to complete thesample separation process. The at least two different motive forces canbe sensitive to timing in that it may be undesirable to activate thesecond motive force until a sufficient volume of sample has been meteredinto one or more of the pathways or until a sufficient fill allows fordrawing of sample into the container without a meniscus break during thedraw process under the second motive force. Suitable methods, devices,features, indicators, or the like can be found in U.S. patentapplication Ser. No. 61/786,351 filed Mar. 15, 2013, fully incorporatedherein by reference for all purposes. Unified holders for multiplecontainers 660, shipping units, additional pieces forattaching/sliding/integrating the containers 660 and/or their holders tothe sample collection/separation device, frits, and other adapterchannels structures can also be found in U.S. patent application Ser.No. 61/786,351 filed Mar. 15, 2013.

FIG. 60 shows a still further embodiment wherein the output tube,needle, channel, or other structure 670 can be oriented to exit from abottom part of the device. It can be orthogonal to the plane or at otherangles. Some embodiments may have both bottom and side exiting outputstructures 670. Some embodiments may have multiple output structures 670in side and/or bottom surfaces.

In one embodiment, the collection and/or separation pathways such as butnot limited to channels may also have a selected cross-sectional shape.Some embodiments of the pathways may have the same cross-sectional shapealong the entire length of the pathway. Optionally, the cross-sectionalshape may remain the same or may vary along the length. For example,some embodiments may have one shape at one location and a differentshape at one or more different locations along the length of thepathways. Some embodiments may have one pathways with onecross-sectional shape and at least one other pathway of a differentcross-sectional shape. By way of non-limiting example, some may have acircular, elliptical, triangular, quadrilateral (e.g., square,rectangular, trapezoidal), pentagonal, hexagonal, octagonal, or anyother cross-sectional shape. The cross-sectional shape may be the samefor the body, support, and base, or may vary. Some embodiments mayselect a shape to maximize volume of liquid that can be held in thepathways for a specific pathway width and/or height. Some may have oneof the pathways with one cross-sectional shape while another pathway hasa different cross-sectional shape. In one embodiment, thecross-sectional shape of the pathway can help maximize volume therein,but optionally, it can also optimize the capillary pulling forces on theblood. This will allow for maximized rate of filling. It should beunderstood that in some embodiments, the cross-sectional shape of thepathway can directly affect the capillary forces. By way of non-limitingexample, a volume of sample can be contained in a shallow but widepathway, or a rounded pathway, both containing the same volume, but onemight be desirable over the other for filling speed, less possibility ofair entrapment, or factors related the performance of the pathway.

Although the pathways may have any shape or size, some embodiments areconfigured such that the pathway exhibits a capillary action when incontact with sample fluid. In some instances, the pathway may have across-sectional area of less than or equal to about 10 mm², 7 mm², 5mm², 4 mm², 3 mm², 2.5 mm², 2 mm², 1.5 mm², 1 mm², 0.8 mm², 0.5 mm², 0.3mm², or 0.1 mm². The cross-sectional size may remain the same or mayvary along the length. Some embodiments may tailor for greater forcealong a certain length and then less in a different length. Thecross-sectional shape may remain the same or may vary along the length.Some pathways are straight in configuration. Some embodiments may havecurved or other shaped path shapes alone or in combination with straightportions. Some may have different orientations within the device body.For example, when the device is held substantially horizontally, one ormore pathways may slope downward, slope upward, or not slope at all asit carries fluid away from the initial collection point on the device.

In some embodiments the inner surface of the pathway and/or othersurfaces along the fluid pathway such as but not limited to the sampleinlet to the interior of a sample collection vessel may be coated with asurfactant and/or an anti-coagulant solution. The surfactant provides awettable surface to the hydrophobic layers of the fluidic device andfacilitate filling of the metering pathway with the liquid sample, e.g.,blood. The anti-coagulant solution helps prevent the sample, e.g.,blood, from clotting when provided to the fluidic device. Exemplarysurfactants that can be used include without limitation, Tween,TWEEN®20, Thesit®, sodium deoxycholate, Triton, Triton®X-100, Pluronicand/or other non-hemolytic detergents that provide the proper wettingcharacteristics of a surfactant. EDTA and heparin are non-limitinganti-coagulants that can be used. In one non-limiting example, theembodiment the solution comprises 2% Tween, 25 mg/mL EDTA in 50%Methanol/50% H20, which is then air dried. A methanol/water mixtureprovides a means of dissolving the EDTA and Tween, and also driesquickly from the surface of the plastic. The solution can be applied tothe pathway or other surfaces along the fluid flow pathway by anytechnique that will ensure an even film over the surfaces to be coated,such as, e.g., pipetting, spraying, printing, or wicking.

It should also be understood for any of the embodiments herein that acoating in the pathway may extend along the entire path of the pathway.Optionally, the coating may cover a majority but not all of the pathway.Optionally, some embodiments may not cover the pathway in the areasnearest the entry opening to minimize the risk of cross-contamination,wherein coating material from one pathway migrates into nearby pathwaysby way of the pathways all being in contact with the target sample fluidat the same time and thus having a connecting fluid pathway.

Although embodiments herein are shown with two separate pathways in thesample collection device, it should be understood that some embodimentsmay use more than two separate pathways. Optionally, some embodimentsmay use less than two fully separate pathways. Some embodiments may onlyuse one separate pathway. Optionally, some embodiments may use aninverted Y-pathway that starts initially as one pathway and then splitsinto two or more pathways. Any of these concepts may be adapted for usewith other embodiments described herein.

Optionally, one or more of the pathways may be coated with a material tobe incorporated into the sample. Optionally, it is desirable to fill theseparator as quickly as possible relative to the other pathway in orderto allow for maximum pre-filtration via the passive mechanisms describedabove. Thus, in one embodiment, one of the pathways fills first beforethe unfiltered/separated pathway fills. In one embodiment, the samplevolume in one pathway is greater than the sample volume in the otherpathway. In one embodiment, the sample volume in one pathway is greaterby 1× than the sample volume in the other pathway.

Optionally, a cap (not shown for ease of illustration) may attach to thecollection device using any technique known or later developed in theart. For instance, the cap may be snap fit, twist on, friction-fit,clamp on, have magnetic portions, tie in, utilize elastic portions,and/or may removably connect to the collection device body. The cap mayform a fluid-tight seal with the collection device body. The cap may beformed from an opaque, transparent, or translucent material.

Optionally, the collection device body of the sample collection andseparation device may be formed in whole or in part from an opticallytransmissive material. By way of non-limiting example, the collectiondevice body may be formed from a transparent or translucent materialsuch as but not limited to Poly(methyl methacrylate) (PMMA),Polyethylene terephthalate (PET), Polyethylene TerephtalateGlycol-modified (PETG or PET-G), or the like. Optionally, only selectpotions of the body are transparent or translucent to visualize thefluid collection channel(s). Optionally, the body comprises an opaquematerial but an opening and/or a window can be formed in the body toshow fill levels therein. The collection device body may enable a userto view the channels within and/or passing through the device body. Thechannels may be formed of a transparent or translucent material that maypermit a user to see whether sample has traveled through the channels.The channels may have substantially the same length. In some instances asupport may be formed of an opaque material, a transparent material, ora translucent material. The support may or may not have the same opticalcharacteristics of the collection device body. The support may be formedfrom a different material as the collection device body, or from thesame material as the collection device body.

The collection device body may have any shape or size. In some examples,the collection device body may have a circular, elliptical, triangular,quadrilateral (e.g., square, rectangular, trapezoidal), pentagonal,hexagonal, octagonal, or any other cross-sectional shape. Thecross-sectional shape may remain the same or may vary along the lengthof the collection device body. In some instances, the collection devicebody may have a cross-sectional area of less than or equal to about 10cm², 7 cm², 5 cm², 4 cm², 3 cm², 2.5 cm², 2 cm², 1.5 cm², 1 cm², 0.8cm², 0.5 cm², 0.3 cm², or 0.1 cm². The cross-sectional area may vary ormay remain the same along the length of the collection device body 120.The collection device body may have a length of less than or equal toabout 20 cm, 15 cm, 12 cm, 10 cm, 9 cm, 8 cm, 7 cm, 6 cm, 5 cm, 4 cm, 3cm, 2 cm, 1 cm, 0.5 cm, or 0.1 cm. The collection device body may have agreater or lesser length than the cap, support or base, or an equallength to the cap, support, or base. There may be variations andalternatives to the embodiments described herein.

Referring now to FIG. 61, a still further embodiment of a samplecollection and sample separation device will now be described. Thisembodiment shows a cartridge 1400 with a sample collection and sampleseparation device 1402 integrated therein having one or two pathways 700and 702. It should be understood that the device 1402 may be integrallyformed with the cartridge. Optionally, it may be a separate unit thatthis is removable from the cartridge. Optionally, it may be a separateunit that this is added to the cartridge after sample has been collectedfrom the subject. Optionally, it may be a separate unit that this isadded and/or attached to the cartridge and sample is collected from thesubject after the unit it added and/or attached to the cartridge.

In this non-limiting example, there is a collection location 1322 andone or more sample openings 1325 and 1329 where sample collection atlocation 1322 can then be accessed such as but not limited to handlingby a pipette tip (not shown). The sample from droplet D will travelalong pathway 1326 as indicated by arrow towards the openings 1325 and1329, where the sample in the opening and any in the pathways 1324and/or 1326 leading towards their respective openings 1325 and 1329 aredrawn into a sample handling system such as but not limited to a pipetteP. In some embodiments, particularly for the pathway 702 with separationmember and the distributor channel 500, a vacuum or suction by thesample handling device can be used to fully draw sample though theseparator 720 and complete the separation process. As indicated byarrows near the pipette P, the pipette P is movable in at least one axisto enable transport of sample fluid to the desired location(s). Althoughonly a single pipette P is shown in FIG. 61 for ease of illustration, itshould be understood that other embodiments may use a plurality ofpipettes to engage one or more items associated with the cartridge. Inthis embodiment, the cartridge 1400 can have a plurality of holdingcontainers 1410 for reagents, wash fluids, mixing area, incubationareas, or the like. Optionally, some embodiments of the cartridge 1400may not include any holding containers or optionally, only one or twotypes of holding containers. Optionally, in some embodiments, theholding containers may be pipette tips. Optionally, in some embodiments,the holding containers are pipette tips that are treated to containreagent(s) on the tip surface (typically the interior tip surfacealthough other surfaces are not excluded). Optionally, some embodimentsof the cartridge 1400 may include only the sample collection device 1402without the tissue penetrating member or vice versa. A suitable devicefor use with cartridge can be found in U.S. patent application Ser. No.13/769,798, filed Feb. 18, 2013. It should be understood that someembodiments may be configured to have only one of the above pathways inthe sample collection and/or sample separation device. Some may havemore than two of the pathways. Some may have more than one separator perpathway. Some may have multiple pathways each with one or moreseparators. Some embodiments may use the sample handling device such asbut not limited to the pipette P to draw sample towards or onto theseparator and then use the pipette to draw sample out of the undersideor other side of the processor after the sample has been or has begun tobe separated.

It should be understood that other cartridge configuration are notexcluded. Some embodiments may directly incorporate the separator and/ordistributor and/or collector into and integrated as part of thecartridge body.

Referring now to FIG. 62, a perspective view is shown of a still furtherembodiment showing a device such as a fluid circuit portion 800 withextraction ports 802 and 804. The embodiment of FIG. 62 uses a singleinlet 806 to direct portions of the sample to two different pathways,wherein at least one portion passes through a formed componentseparation member.

Referring now to FIG. 63, a perspective view is shown of a still furtherembodiment showing a device with fluidic circuit portion 800, a housingportion 822, and a sample container unit 824. As seen in FIG. 63, thisnon-limiting sample shows housing portion 822 coupling the fluidiccircuit portion 800 with the sample container unit 824. The housingportion 822 allows for the sample container unit 824 to be coupled tothe same fluidic circuit portion 800 but still have the sample containerunit 824 movable between a first position (as shown in FIG. 63) and asecond position.

In one embodiment, an inlet port centered along a mid-line of thefluidic circuit with an entry point on to the formed componentseparation member that is off-center relative to the midline of theformed component separation member.

In one embodiment, a vent channel of a curved configuration is coupledto a vent channel with a curved portion and an intersection linear (bentor straight) portion. The vents, in this non-limiting example arecollection features that are on the side of the separation member werefluid, but not formed components, can exit the membrane. In onembodiment, a vent channel has a linear (bent or straight) portioncoupled to a distribution portion, wherein the linear portion is closerto an external vent and distribution portion is closer to the separator.

Referring now to FIGS. 64 to 67, various embodiments of thecross-sectional shapes of the capillary structure. FIG. 64 showsstructures with sharp corners 890 while FIG. 65 shows an embodiment withradii or rounded corners 892.

As more clearly seen in the non-limiting example of FIG. 67A, tangencyand curvature of the rounded corners 892 may provide continuous liquidcontact to assist with fluid flow out of a separation member such as butnot limited to a membrane and into an opposing portion of the fluidcircuit such as but not limited to the capillary flow structure. Itshould be understood that the round corners 892 creates at least onetransition region 894 which can assist drawing or leaching of fluid fromone region towards the fluid collection structures in the device. Insome embodiments, this drawing or leaching of fluid out of theseparation member can be desirable. By contrast, the embodiments with asharp corner 890 which opens directly the capillary structure with atransition region diminishes the assistance that may come from have theclosely spaced area associated with the tangency and curvature providedby rounded corners 892. Optionally as seen in FIG. 67B, some embodimentsmay have a sharp corner 896 but further include at least one transitionregion that may provide continuous liquid contact to assist with fluidflow out of the separation member. Optionally, some embodiments may haveat least one increased width region between the capillary structure 898and the transition region 894.

Optionally in one non-limiting example, the capillary structure may beformed of or have a surface treated to create a hydrophilic fluidicstructure. In one non-limiting example, the structure can be made of ahydrophilic material such as but not limited Polyethylene terephthalateglycol-modified (PET-G) which has a small wetting angle and is ahydrophilic material which can draw fluid toward the back side of themembrane. Optionally, some embodiments may use cellulose acetate,cellulose acetate butyrate, or other suitable material.

Plasma collection vent: more plasma without hemolysis; distributors alsoare; capillary structures on the back side membrane. The other vent(blood side/distributor vents) is shown on the top side of theseparation device that coupled to vent 915. Alternate plasma extractionmethods are provided wherein different motive force, other than vacuumin a container, are used to draw fluid away from the separationmembrane. In one embodiment, inlet flow control features may be used inthe sample container to control the rate and/or amount of motive forceapplied to filtered sample and/or sample about to be filtered. It shouldbe understood that hemolysis will corrupt the sample for many assays andis thus generally undesirable. Optionally, some embodiments may gowithout a dual channel inlet and use a single channel. Some embodimentsmay have an opening over the membrane, instead of at one end.

Optionally, this embodiment can have a passive, always open vent insteadof a valve.

Optionally, some alternate extraction methods may include: providing amuch higher extraction vacuum (crimp opening to meter pull forces,wherein the crimp results in a 5 to 10 micron wide opening in the tube,almost a cold weld when cutting so as to form a flow regulator). Itshould be understood that high vacuum in the container or from wasanother source was high enough to collect a desired liquid volume, butthe initial spike from the high vacuum will cause excess pull on theform components that creates hemolysis when the sample is a bloodsample. In one embodiment, there is at least 70% of theoretical fluidrecovery. In one embodiment, there is at least 80% of theoretical fluidrecovery. In one embodiment, there is at least 90% of theoretical fluidrecovery.

For the final steps, the amount of friction can provide sufficientmechanical resistance from rapidly pushing sample vessel rapidly intothe holder. The friction can be from the plunger, an external guide,and/or other component to provide a controlled movement between a firstposition and a second position. Other mechanical mechanism can be usedto regulate speed that the user pushes it on. On the non-separator side,in one non-limiting example, there is no plunger. One embodiment may usedeflected point needle that is anti-coring and also provides a sideopening needle tip. In one-nonlimiting example, the anti-coring isdesirable to prevent coring of the frit, which may introduce undesirablefrit parts into the sample. The needle pierces through the frit. Itshould be understood that the frit is sized to cover or at leastsubstantially cover the opening of the needle pointed tip opening.

In this non-limiting example, the plunger may have a harder portion inthe center while a circumferential portion is softer for liquid sealperformance.

Optionally, the capillary channels with fluid therein can also settle abit before being engaged to be extracted. This delayed fill of thenon-separator side ensures that the separator side has filled and hadsome settling time before being engaged to the sample collection unitfor fluid transfer into the sample collection unit. In one embodiment,80 microliters of whole blood results in 16 to 20 microliters of plasma.

Referring now to FIG. 68, a cross-sectional view is shown of onenon-limiting example wherein an inlet channel 808 is shown penetratingone non-limiting example of a sample container unit 824. As seen in FIG.68, the movement of the plunger 828 of the sample container unit 824 canbe used to create a motive force such as but not limited to at least apartial vacuum to draw liquid from the channel 808 into the samplecontainer unit 824. In this non-limiting example, as the plunger isdisplaced as shown by arrow 831 in FIG. 68, this increases the interiorvolume 829 of the sample container unit 824 between the cap portion 832.It should be understood that, in one non-limiting example, there may beno sample in the sample container unit 824 until the motive force isprovided to overcome any inherent capillary force of the channel 808which brings the sample fluid into but not out a needle end 834 of thechannel 808, In one non-limiting example, extracting fluid from thechannel 808 may involve using one or more additional motive forces. Itshould be understood that this configured described herein may besimilar to a reverse plunger. Optionally, some embodiments may use aconventional plunger, in place of or in combination with the structuresherein, to provide motive force to draw sample into the samplecontainer.

FIG. 68 also shows that, in at least one embodiment, the channel 808 mayhave a pointed distal end 834. Still further embodiments may have thechannel 808 be of a non-coring design so as not to introduce any coredportion or debris of the cap 832 into the collected fluid. Regardless ofwhether a non-coring, conventional, or other shaped channel 808, itshould be understood that some embodiments of plunger 828 may have ahardened core portion 838 that can withstand force input from thechannel 808. As seen in FIG. 68, at least some embodiments will have acompliant material between the hardened core portion 838 and the sidewalls of the sample container so as to provide at least a sufficientfluid seal as the plunger 828 is moved from at least a first position toat least a second position.

Referring now to the non-limiting examples of FIGS. 69 and 70, inletdesign of inlet 806 may include flow guide structures 910 and 912 suchas one or more small capillary channels in the sides of the inlet 806encourage flow to enter pathway leading to the separation member, ratherthan the pathway without the separation member. As seen in FIG. 73, theflow guide structures 910 and 912 are located as positions along theinlet 806 that have surfaces that extend toward the separation component920. In one non-limiting example, the fluid guide structures 910 and 912are not along the bottom surface of the inlet, which may be where theother channel connects to the inlet. In one non-limiting example, thefluid guide structures 910 and 912 are not positioned along a surface ofthe inlet where the other channel connects to the inlet. Although FIG.69 shows that the inlet 806 has at least two flow guide structures 910and 912, it should be understood that some embodiments may only have asingle flow guide structure. Optionally, some embodiments may have morethan two flow guide structures. Optionally, some embodiments may have asingle structure at the inlet 806 but forks into two or more structuresas the fluid flows deeper into the inlet. Optionally, some embodimentsmay have a plurality of fluid guide structures wherein at least two ofthe structures merge together so that there are fewer guide structuresas the fluid flow deeper into the structure. It should be understoodthat some embodiments may take a plurality of guide structures and mergethem all into one guide structure.

FIG. 71 shows a still further embodiment wherein at least one stopstructure 930 such as but not limited to a frit is included on at leastone of the extraction channels 808. In the non-limiting example of FIG.71, the stop structure 930 is included on the channel 808 coupled to thenon-separation member pathway, which may flow more freely and thus havea different flow resistance than the other pathway which passes throughthe separation member. Venting of the non-separation member pathwaychannel allows filling via capillary flow. In this embodiment, thechannel 808 is vented through a pointed end of a needle, wherein an airporous frit such as but not limited to one of Porex or similar porousmaterial, is coupled to a tip of needle, still allowing air through, butwith more resistance. In this manner, filling on the non-separationmember pathway side is slowed down so that separation member pathway canfill first. Other techniques for slowing flow along one pathway are notexcluded and may be used alone or in combination with the stop structure930 discussed herein. FIG. 71 also shows that in this embodiment, acombined vent 915 can be used to provide a vent path for the variousvents associated with the sample distributor over the fluid entrysurface of the separation member.

Furthermore, fill metering can be done using an indicator 950 (see FIG.73) on the non-separation member pathway, because due to its laggingindicator quality due to a slower fill, if a fill level is reached onthe non-separation member pathway, due to the slower, fill a user cansafely conclude that the other pathway has already completed its fillprocess due to the slower fill in the non-separation member pathway.FIG. 73 also shows that there are at least two fluid flow paths withinthe device as indicated by arrows 951 and 953. FIG. 73 also shows thatthere may be guide 955, such as but not limited a guide member in aslot, that may act as a visual indicator that the movement of the samplecollection unit 824 is complete and may optionally provide sufficientresistance to encourage a controlled rate of movement of the collectionunit 824 so that the flow will be sufficient to minimize hemolysis offormed components in the sample.

Referring now to FIG. 72A, it should also be understood that structureswithout a penetration tip, such a pipette tip, can also be adapted foruse with certain embodiments of the sample container. FIG. 72A alsoshows that in at least some embodiments, the plunger 828 is only in onevessel and not in all of the vessels defined by the sample containerunit. FIG. 72A may also be shown with the top plug 832 removed to allowfor sample extraction using a pipette tip that is shaped to reach abottom interior portion of the vessel being emptied.

Referring now to the non-limiting example of FIG. 72B, the variousinitial positions are shown for a stop structure 930 on the channel 808and that non-coring tip of the channel 808 has engaged a plug 832 of thecontainer unit 824.

Referring now to FIG. 74, this non-limiting example shows a samplecontainer unit 824 with one sample container sized larger to accommodatea pressure drop volume, that is the volume to which air on the plasmaside of the formed member separation membrane (including inside thesample container unit) expands due to the pressure drop across themembrane.

FIG. 75 shows a side view of the device wherein the sample flow pathwayindicated by arrow 1010 shows that sample enters at an angle, flowsalong one plane, downward to a different plane and is drawn laterallyout at the lower plane. Movement of the sample collection unit 824 isindicated by arrow 1020, wherein in this embodiment, movement of thesample collection unit 824 provides motive force to draw samplesubstantially free of formed components into the sample collection unit.Although many of the embodiments shown herein use linear movement of thesample collection unit 824, it should be understood that embodimentsusing rotary motion to provide the motive force or rotary motiontranslated into linear motion to provide the desired movement to drawsample into the sample container.

Some embodiments of the collection unit may have a cross-sectional shapewith an asymmetry, a protrusion, or other feature that serves as akeying feature for orienting the SCU in any receiving device orstructure.

While the invention has been described and illustrated with reference tocertain particular embodiments thereof, those skilled in the art willappreciate that various adaptations, changes, modifications,substitutions, deletions, or additions of procedures and protocols maybe made without departing from the spirit and scope of the invention.For example, with any of the above embodiments, it should be understoodthat some embodiments may handle other types of samples and necessarilybiological samples. Although many illustrations are shown with only asingle inlet port, it should be understood that some embodiments mayhave at least two inlet ports. In some embodiments, both inlet ports areon the same end of the device. Optionally, some embodiments may haveinlet ports on the same surface of the device. Optionally, at least thetwo inlets are adjacent to each other. Optionally, there are at leastthree inlet ports. Optionally, at least two inlet ports are each definedby at least one capillary tube. In this embodiment where each inlet hasits own capillary tube, at least one tube directs fluid to anon-separation pathway while a second tube directs fluid to a separationpathway. Optionally, some embodiments may combine inlets formed bycapillary tubes with inlet(s) associated with a non-capillary pathway.Some embodiments may have the inlet along a centerline axis of thedevice. Optionally, some embodiments may have the inlet aligned off thecenterline. Optionally, some embodiments may orient the inlet to bealong or parallel to the axis of the centerline of the device.Optionally, some embodiments may orient the inlet along an axis that isat an angle to the plane of the device. Optionally, instead of havingthe inlet at one end of the separation device, it should be understoodthat some embodiments may have the inlet directly over at least oneportion of the separation device. In this manner, the opening may directfluid onto the membrane with a minimal amount of travel in a lateraltube or pathway.

Optionally, some embodiments may be configured with a co-axial designsuch as shown in FIG. 76. One embodiment may have sample enter along aninner lumen for an inside-out type filtration as indicated by arrow1010. Optionally, some embodiments may use an outside-in type filtrationif the separation membrane is located in the inner lumen and samplefluid enters from a surface opening (shown in phantom) as indicated byarrow 1012 or from an inlet on one end of the device.

Optionally as seen in FIG. 77A, some embodiments may have a portion 800such as the fluid circuit portion that includes the separation memberfluidly coupled to a second portion 1040. Optionally, it may beconfigured not to include the non-separation pathway. Optionally, it maybe configured to include the non-separation pathway. Some embodimentsmay have this combination of portion 800 with portion 1040 in a teststrip configuration. Some embodiments may have this combination ofportion 800 with portion 1040 in a lateral flow device configuration.Some embodiments may have a unibody structure or other merged structurethat is formed to provide support to both portions 800 and 1040. Motiveforce can be provided to move the sample as indicated by arrow 1030which flow out or the fluid circuit of portion 800 and that the fluidportion of the sample, substantially free of the formed components, canenter a second region 1040 which may be but is not limited to ananalytical region. In some embodiments, the second region 1040 alsoprovides a motive force such as but not limited to wicking forceassociated with such material in at least a portion of the second region1040.

FIG. 77B shows a still further embodiment wherein the sample collectiondevice has a plurality of tissue penetrating member or members 1292mounted to an actuation mechanism 1293. In one embodiment, the tissuepenetrating members 1292 are microneedles. In one embodiment, the tissuepenetrating member 1292 comprises a lancet. In one embodiment, theactuation mechanism 1293 can be a spring-like device in a dome, curved,or other shape. In some embodiments, the dome shape can also provide acertain suction force to draw sample upward from the collection site.Although a mechanical actuation method is shown, it should be understoodthat other types of actuation techniques such as but not limited toelectromechanical, pneumatic, mechanical cam, or other technique knownor developed in the future may be used for actuation. It should also beunderstood that some embodiments may use a tissue interface (shown inphantom) to facilitate interaction with the tissue. FIG. 77B shows thatthe sample obtained from a wound or wounds created by tissue penetratingmembers 1292 may flow through channel(s), capillary tube(s), or otherpathways as indicated by arrow 1295 to a channel 1299 or other inlet toa separation device. In one embodiment, the channel 1299 may be coatedwith at least one anticoagulant. Optionally, some embodiments may havetwo channels 1299 that may draw sample along two pathways, wherein eachchannel may have the same, or optionally different, coatings on thesurface of the channels 1299. Optionally, some embodiments may havesurfaces of the device uncoated but instead have the additive materialin the container 1297.

As seen in FIG. 77B, the sample may flow to a location such as but notlimited to a chamber (shown in phantom), one end of the channel 1299 orother location wherein a conduit such as pathway 1301 or 1303 (shown inphantom) may be used to fluidically couple the sample collected inchannel 1299 to transfer the sample container 1296 or 1297. Motive forcecan be provided to move the sample as indicated by arrow 1030 along ahorizontal path over the separator, a path from the plane of above theseparator to a plane below the separator, and then laterally towards atleast one sample container. In this non-limiting example, the pathway isa zig-zag path from one side of the separator to the other side whichcarries the liquid portion to an intermediate chamber or directly to oneor more containers. As described, the container may be one with a subatmospheric condition therein (prior to being fluidically engaged to thefluid pathway), one with a reverse syringe design as found in U.S.Provisional Application Ser. No. 62/051,906 filed Sep. 17, 2014, acontainer may be one that is un-pressurized, without a movable plungerbut with a septa cap, or other suitable container for sample ascurrently known or may be developed in the future.

It should be understood that some embodiments may have containers 1296in both locations as shown in FIG. 77B or only at one but not the other.Optionally, some may have multiple containers at one location and noneor fewer containers at the other location. Optionally, some of these maybe unitized so that multiple vessels are integrally formed or otherwisejoined together. As seen in FIG. 77B, the container 1296 may be actuatedby sliding the container 1296 to contact the pathway 1301 in a mannerthat allows sub-atmospheric environment inside the container 1296 todraw sample therein. Optionally, other actuation methods such as but notlimited to using a valve, breaking a seal, or the like can be used toactivate sample transfer from the device to the container 1296. Someembodiments may keep the channel 1299 in one horizontal plane or mayoptionally have portions in one plane and portions in another plane.Optionally, instead of or in combination with capillary action fromchannel 1299 for drawing sample therein from the wound site, a suctionor other sample pulling device can be used to draw sample into thechannel 1299. The embodiment of FIG. 77B may optionally be modified tolocate the entry port of channel 1299 closer to the wound site such asbut not limited a channel extension 1305, forming the channel closer tothe wound site, or positioning or orienting the tissue penetrationmembers to form a wound closer to the inlet of channel 1299. It shouldbe understood that devices herein may be configured to include featuresfrom U.S. Provisional Application Ser. No. 62/051,906 filed Sep. 17,2014, fully incorporated herein by reference for all purposes. It shouldbe understood that devices in U.S. Provisional Application Ser. No.62/051,906 filed Sep. 17, 2014, may be configured to include a formedcomponent separation apparatus as described in this application.

FIG. 77B also shows a still further embodiment using container 1297having a reverse-syringe design is used. As seen herein, the movement ofengaging the container 1297 with pathway 1301 or 1303 can be used topush the plunger 2828 to create a reduced pressure environment thatdraws sample into the container 1297. It should be understood that someembodiments may have containers 1297 in both locations as shown in FIG.77B or only at one but not the other. Optionally, some may have multiplecontainers at one location and none or fewer containers at the otherlocation. Optionally, some of these may be unitized so that multiplevessels are integrally formed or otherwise joined together. Optionally,some embodiments may have one type of container 1296 at one location anda different type of container 1297 at a different location shown in FIG.77B. Optionally, some may have at least two different types of containerat one location. Referring still to FIG. 77B, some embodiments may use apush element 1307 that provide a cap or other seal that when moved asindicated over feature 1305 will cause a pressurized air bolus to pushsample in the channel 1299 outward into the containers 1297 that may beattached to 1301 or 1303.

Optionally, it should be understood that some embodiments may have atleast one formed component separation pathway for use in anon-diagnostic device. By way of non-limiting example, the device may befor sample collection, where no diagnosis occur on the device.Optionally, it should be understood that some embodiments may have atleast one formed component separation pathway and at least onenon-separation pathway for use in a non-diagnostic device. Optionally,it should be understood that some embodiments may have at least twoformed component separation pathway and at least one non-separationpathway for use in a non-diagnostic device. Optionally, it should beunderstood that some embodiments may have at least one formed componentseparation pathway and at least two non-separation pathways, all for usein a non-diagnostic device. Of course, some alternative embodiments mayhave one or more pathways for use for diagnosis. Optionally, someembodiments may use this type of separation device with lateral flowstrip wherein the fluid, after formed component separation, may be movedsuch as but not limited to wicking or other capillary flow onto a secondregion such as but not limited to an analyte-detecting region on adevice such as but not limited to a test strip for analysis.

Optionally, some embodiments may provide a vibration motion source, suchas but not limited to one built into the device and/or in an externaldevice use to process the sample container, to assist in fluid flowwithin device, during the collection, or post-collection. Someembodiments may use this vibration to assist flow or to remove any airpockets that may be created, such as but not limited to when doing atop-down fill. Optionally, some embodiments may provide more periodic orpulse type force to assist in fluid flow.

It should be understood that although many components herein are shownto be in alignment in the same plane or parallel planes, someembodiments may be configured to have one or more component in a planeangled to or orthogonal to a plane of the fluid collection circuit inportion 800. The fluid collection circuit in portion 800 does not needto be a flat planar device and may be in a curved configuration.Optionally, some embodiments may have it a cone configuration.Optionally, some embodiments may have it device with a polygonalcross-sectional shape. As seen, the fluid collection circuit in portion800 is not limited to a planar shape.

It should also be understood that in many embodiments, the portion 800may be made of a transparent material. Optionally, the portion 800 maybe made of a translucent material. Optionally, portions of the portion800 may be covered with paint or other opaque material, be formed of anopaque material, or the like such that only portions that may containfluid are transparent or translucent so as to provide an indicator offill level. Such an embodiment may have all or only a portion of thefluidpath visible to the user. In one non-limiting example, bar codes,color-coding, visual information, instructions, instructions for use,fill-indicator, advertising, child-appealing aesthetics, texturing,texturing for grip purpose, texturing for contour, texturing to providefeedback such as orientation of the front of the device, or othercoating may be used hereon.

Optionally, some embodiments may include an intermediary structurebetween the fluid circuit in portion 800 and the sample collection unit824. This intermediary structure can be in the fluid pathway and providecertain function such as but not limited to introducing a material intothe collected fluid such as but not limited to anti-coagulant.Optionally, the intermediary structure in the fluid path may provideanother route, such as switch or connection pathway, to add additionalsample or other liquid material into the collected fluid.

Optionally, some embodiments may have disposable portion(s) and reusableportions, wherein the reusable portions can be mated with the disposableportion(s) to form another collection device. By way of non-limitingexample, a reusable portion may be one that does not directly contactthe sample fluid or filtered fluid.

Although embodiments herein show the separation member as part of ahandheld device, it should be understood that other embodiments mayincorporate the device as part of a non-handheld benchtop device, anon-portable device, or the like and the disclosures herein are notlimited to handheld or disposable units. Some embodiments may alsoinclude features for collection sample from a plurality of sampleprocessing devices. In this manner, an increased amount of filteredsample can be collected, simply by using more devices for use with moresamples which in one embodiment may all be from one subject. Optionally,samples in multiple devices may be from multiple subjects.

As used herein, the terms “substantial” means more than a minimal orinsignificant amount; and “substantially” means more than a minimally orinsignificantly. Thus, for example, the phrase “substantiallydifferent”, as used herein, denotes a sufficiently high degree ofdifference between two numeric values such that one of skill in the artwould consider the difference between the two values to be ofstatistical significance within the context of the characteristicmeasured by said values. Thus, the difference between two values thatare substantially different from each other is typically greater thanabout 10%, and may be greater than about 20%, preferably greater thanabout 30%, preferably greater than about 40%, preferably greater thanabout 50% as a function of the reference value or comparator value.

As used herein, a “surfactant” is a compound effective to reduce thesurface tension of a liquid, such as water. A surfactant is typically anamphiphilic compound, possessing both hydrophilic and hydrophobicproperties, and may be effective to aid in the solubilization of othercompounds. A surfactant may be, e.g., a hydrophilic surfactant, alipophilic surfactant, or other compound, or mixtures thereof. Somesurfactants comprise salts of long-chain aliphatic bases or acids, orhydrophilic moieties such as sugars. Surfactants include anionic,cationic, zwitterionic, and non-ionic compounds (where the term“non-ionic” refers to a molecule that does not ionize in solution, i.e.,is “ionically” inert). For example, surfactants useful in the reagents,assays, methods, kits, and for use in the devices and systems disclosedherein include, for example, Tergitol™ nonionic surfactants and Dowfax™anionic surfactants (Dow Chemical Company, Midland, Mich. 48642);polysorbates (polyoxyethylenesorbitans), e.g., polysorbate 20,polysorbate 80, e.g., sold as TWEEN® surfactants (ICI Americas, NewJersey, 08807); poloxamers (e.g., ethylene oxide/propylene oxide blockcopolymers) such as Pluronics® compounds (BASF, Florham Park, N.J.);polyethylene glycols and derivatives thereof, including Triton'surfactants (e.g., Triton' X-100; Dow Chemical Company, Midland, Mich.48642) and other polyethylene glycols, including PEG-10 laurate, PEG-12laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate,PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate,PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryltrioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryllaurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate,PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castoroil, PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castoroil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol, PEG-30soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitanlaurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearylether, tocopheryl PEG-100 succinate, PEG-24 cholesterol,polyglyceryl-10oleate, sucrose monostearate, sucrose monolaurate,sucrose monopalmitate, PEG 10-100 nonyl phenol series, PEG 15-100 octylphenol series, and poloxamers; polyoxyalkylene alkyl ethers such aspolyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such aspolyethylene glycol alkyl phenols; polyoxyalkylene alkyl phenol fattyacid esters such as polyethylene glycol fatty acids monoesters andpolyethylene glycol fatty acids diesters; polyethylene glycol glycerolfatty acid esters; polyglycerol fatty acid esters; polyoxyalkylenesorbitan fatty acid esters such as polyethylene glycol sorbitan fattyacid esters; phosphocholines, such as n-dodecylphosphocholine, (DDPC);sodium dodecyl sulfate (SDS); n-lauryl sarcosine;n-dodecyl-N,N-dimethylamine-N-oxide (LADO); n-dodecyl-P-D-maltoside(DDM); decyl maltoside (DM), n-dodecyl-N,N-dimethylamine N-oxide (LADO);n-decyl-N,N-dimethylamine-N-oxide,1,2-diheptanoyl-sn-glycero-3-phosphocholine (DHPC);1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC); 2-methacryloyloxyethylphosphorylcholine (MPC);1-oleoyl-2-hydroxy-sn-glycero-3-[phospho-RAC-(1-glycerol)] (LOPC);1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-RAC-(1-glycerol)] (LLPG);3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS);n-Octyl-N,N-dimethyl-3-ammonio-1-propanesulfonate;n-Decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate;n-Dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate;n-Hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate;Tetradecanoylamidopropyl-dimethylammonio-propanesulfonate;Hexadedecanoylamidopropyl-dimethylammonio-propanesulfonate;4-n-Octylbenzoylamido-propyl-dimethylammonio Sulfobetaine; a Poly(maleicanhydride-alt-1-tetradecene), 3-(dimethylamino)-1-propylaminederivative; a nonyl phenoxylpolyethoxylethanol (NP40) surfactant;alkylammonium salts; fusidic acid salts; fatty acid derivatives of aminoacids, oligopeptides, and polypeptides; glyceride derivatives of aminoacids, oligopeptides, and polypeptides; lecithins and hydrogenatedlecithins, including lecithin, lysolecithin, phosphatidylcholine,phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid,phosphatidylserine; lysolecithins and hydrogenated lysolecithins;phospholipids and derivatives thereof; lysophospholipids and derivativesthereof, including lysophosphatidylcholine,lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidicacid, lysophosphatidylserine, PEG-phosphatidylethanolamine,PVP-phosphatidylethanolamine; carnitine fatty acid ester salts; salts ofalkylsulfates; fatty acid salts; sodium docusate; acyl lactylates; mono-and di-acetylated tartaric acid esters of mono- and di-glycerides;succinylated mono- and di-glycerides; citric acid esters of mono- anddi-glycerides; lactylic esters of fatty acids, stearoyl-2-lactylate,stearoyl lactylate, succinylated monoglycerides, mono/diacetylatedtartaric acid esters of mono/diglycerides, citric acid esters ofmono/diglycerides, cholylsarcosine, caproate, caprylate, caprate,laurate, myristate, palmitate, oleate, ricinoleate, linoleate,linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate,lauroyl carnitines, palmitoyl carnitines, myristoyl carnitines, andsalts and mixtures thereof; alkylmaltosides; alkylthioglucosides; laurylmacrogolglycerides; hydrophilic transesterification products of a polyolwith at least one member of the group consisting of glycerides,vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols;polyoxyethylene sterols, derivatives, and analogues thereof;polyoxyethylated vitamins and derivatives thereof;polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof;fatty alcohols; glycerol fatty acid esters; acetylated glycerol fattyacid esters; lower alcohol fatty acids esters; propylene glycol fattyacid esters; sorbitan fatty acid esters; polyethylene glycol sorbitanfatty acid esters; sterols and sterol derivatives; polyoxyethylatedsterols and sterol derivatives; polyethylene glycol alkyl ethers; sugaresters; sugar ethers; lactic acid derivatives of mono- anddi-glycerides; hydrophobic transesterification products of a polyol withat least one member of the group consisting of glycerides, vegetableoils, hydrogenated vegetable oils, fatty acids and sterols; oil-solublevitamins/vitamin derivatives; and combinations thereof

Referring now to FIG. 78, one embodiment of a filtering device such asbut not limited to a bodily fluid separation material 3100 will now bedescribed. FIG. 78 shows a side cross-sectional view of the separationmaterial 3100, showing cross-sections of the structures 3102 of theseparation material. By way of non-limiting example, the separationmaterial 3100 may be a size-exclusion barrier such as but not limited toa porous membrane with size-exclusion properties. Other embodiments mayuse other types of size-exclusion barrier(s). In one embodimentdescribed herein, the structures 3102 are fibers in the separationmaterial with their cross-sectional views shown in FIG. 78. Optionally,the structures 3102 are mesh portions of the separation material.Optionally, the structures 3102 are pore walls or pore-definingstructures of the separation material. Optionally, the structures 3102may be a percolating network of connected fibers, elongate members, orthe like. Some embodiments may combine one or more of the foregoing toform the separation material. Although the descriptions herein arewritten in the context of a separation material, other filter materialsor structures in sheet-like or other shapes are not excluded material.FIG. 78 shows that for the present embodiment, formed components 3106such as but not limited to red blood cells, white blood cells, platelet,or other formed components of the bodily fluid can enter the separationmaterial 3100 in a variety of directions, including from a top-downmanner, and will continue to pass through the separation material untilthe component reaches a size-constrained area where the spacing becomestoo small for the formed component 3106 to proceed any further. In thisembodiment, operating under the principle of size exclusion, the formedcomponent 3106 will then be constrained in the separation material 3100while liquid portions and/or those components not size excluded cancontinue to pass through the separation material. In one non-limitingexample, arrows 3104 show movement of formed components through theseparation material 3100 of FIG. 78. Other movement, such as but notlimited to lateral, side-ways, and/or diagonal movement, is notexcluded.

Referring still to the embodiment of FIG. 78, the dotted line 3120 showsthat in this embodiment, there are at least two regions 3122 and 3124for the separation material 3100. It should be understood that otherembodiments can have even more regions. In this current embodiment, theregion 3122 comprises a formed component capture region. In somespecific embodiments as will be discussed in more detail below, it maybe an anti-hemolytic, formed component capture region. By waynon-limiting example, the region 3124 comprises a pass-through regionthat has structural elements spaced closely enough that formedcomponents of the bodily fluid sample cannot completely pass throughthat region 3124. In at least some embodiments, the sizing and/orspacing of elements is selected such that the size-restriction techniqueof separation material components prevents the formed components fromcontinuing through the separation material. This filters out the formedcomponents from the liquid components of the bodily fluid.

In one embodiment, because region 3122 can be configured to be a formedcomponent capture region, structures in the region 3122 will have morepotential direct contact with the formed components 3106 and be incontact with them for a longer period of time, relative to structures inthe second region 3124. Due at least in part to the greater directcontact physically and temporally, it may be desirable in it at leastsome embodiments described herein to treat the structures 3102 of theregion 3122 to minimize undesirable breakdown, spoilage, or otherdetrimental effect that may result from the formed components beingcaptured in the region 3122. In one non-limiting example, the structures3102 may be coated with an anti-hemolytic coating to prevent breakdownof red blood cell when the bodily fluid being processed is blood. Oneembodiment of an anti-hemolytic coating may be an NTA coating.Optionally, other anti-hemolytic treatments in layer or other form mayuse material such as but not limited to n-Octyl-r3-D-Glucopyranoside(OG), cell lipid bilayer intercalating material, phosphate estercontaining at least two ester linkages comprising fatty hydrocarbongroups, tri-2-ethylhexylphosphate, di-2-ethylhexylphthalate,dioctylterephthalate, anti-hemolytic surfactant(s), a surfactant such asbut not limited to polysorbate 80 mixed with any of the foregoing,and/or other anti-hemolytic material. Other anti-hemolytic material usedwith embodiments herein includes but is not limited to one or more ofthe following: anti-coagulants, proteins (such as but not limited toBSA, HSA, Heparin, Casein, etc.), surfactants (such as but not limitedto Tween, Silwet, SDS, etc.), sugars (such as but not limited tosucrose, trealose, etc.), and/or the like.

In one embodiment, the region 3124 may be configured to be a liquidpass-through region positioned after the bodily fluid has passed throughregion 3122. Although FIG. 78 illustrates region 3124 to be next toregion 3122, it should be understood that embodiments havingintermediate region(s) and/or space between the regions are notexcluded. By way of non-limiting example, the pass-through region 3124may be configured not have direct contact with the formed components.Optionally, only structures 3108 defining part of the upper portion ofthe region 3124 may be in contact with any formed components 3106.Optionally, only structures 3108 defining part of the upper surface ofthe region 3124 may be in contact with any formed components 3106. Inone embodiment, the region 3124 may be have a selected structure size,spacing, and/or other property that prevents formed components 3106 frompassing through the region 3124 so as to enable a size restrictionfiltering technique for removing formed components from the bodilysample.

In at least some embodiments, because the formed components are not indirect contact with the region 3124 or are only in minimal contact withregion 3124, the separation material of region 3124 may not be coatedwith the material used in the region 3122. Optionally, region 3124 maybe selective coated with the materials used in region 3122 in a mannersuch as but not limited to only those portions that might still be incontact with formed components may be coated, which others portions ofregion 3122 are uncoated. Optionally, at least some embodiments may havesome or all of region 3124 coated with a material different from that ofthe region 3122. Optionally, at least some embodiments may have some orall of region 3124 covered with the material of region 3122 and thenadding a second layer of the second material over the material of region3122. In one non-limiting example, this second material may be selectedto prevent the first material leaching or otherwise entering the bodilyfluid when the liquid passes through the region 3124. In at least someembodiments, the portions of region 3124 covered with the material ofregion 3122 is covered with the second material while other areas ofregion 3124 are substantially or at least partially uncovered by eithermaterial. By way of example and not limitation, some embodiments may useHeparin and/or other anti-coagulant as the material for the secondlayer. Optionally, the material for the second layer may be a materialthat is already in the bodily fluid sample. By way of non-limitingexample, the material may be EDTA if the bodily fluid sample has alreadybeen or will be treated with EDTA. Optionally, for the second layer,some embodiments may use inert materials alone or in combination withany of the other materials listed herein.

Referring now to FIG. 79, a still further embodiment will now bedescribed. This embodiment shows a first separation material 3200 and asecond separation material 3210. Although only two separation materialsare shown, it should be understood that other embodiments havingadditional separation materials above, between, and/or below theseparation materials shown in FIG. 79 are not excluded. It should alsobe understood that one or more of the separation materials 3200 and 3210can, within the separation materials themselves, each have additionalregions therein for different properties.

As seen in the embodiment of FIG. 79, the separation material 200functions as a capture region similar to the capture region 3122 of theembodiment of FIG. 78. In the current embodiment, the separationmaterial 3210 functions as a pass-through region similar to region 3124of the embodiment of FIG. 78.

Referring now to FIG. 80, this embodiment shows a tri-layer filterassembly with a first layer 3300, a second layer 3310, and a third layer3320. For ease of illustration, the layers are shown to be similar inthickness, but configurations where all three are of differentthicknesses, or only are of different thicknesses are not excluded.Embodiments with additional layers are also not excluded. Layers canalso be formed of different materials.

It should be understood that any of the layers 3300, 3310, or 3320 canbe configured as a capture region, a pass-through region, or neither. Inone non-limiting example, at least the upper two layers 3300 and 3310are capture regions. They can have similar capture capabilities, oroptionally, one can be configured to be preferential capture ofcomponents while the other layer has preferential capture of componentsin a different size and/or shape regime. In another non-limitingexample, at least the upper two layers 3300 and 3310 are capturesregions, but only one of them is coated with a material to preventdegradation of the formed component(s). Optionally, both of them arecoated with a material to prevent degradation of the formedcomponent(s). Another embodiment may have two layers such as layers 3310and 3320 that are both configured as pass-through layers. In oneembodiment, neither of the layers 3310 or 3320 have structures that arecoated with a material to prevent degradation of the formedcomponent(s). Optionally, at least one of the layers 3310 or 3320 hasstructures that are coated with a material to prevent degradation of theformed component(s). Optionally, some embodiments have both of thelayers 3310 or 3320 have structures that are coated with a material toprevent degradation of the formed component(s).

Separation Material Treatment

By way of example and not limitation, in order to be able to useseparation materials for producing plasma suitable for a greater rangeof assays, several separation material treatment methods have beenidentified. Some of these techniques may involve treatment of separationmaterials after they are formed. Some of the techniques may involveforming the separation materials in a way that does not involveadditional treatment after separation material formation. Optionally,some techniques may use both separation material formation andpost-formation treatment to create a desired configuration.

1. Separation material wash: In one embodiment described herein, bycontrolled washing of the coated plasma separation material by waterand/or buffer solutions, most of the hemolysis-preventing agents can beremoved. FIG. 81 shows that a washing mechanism, such as but not limitedto a nozzle 3400 directing washing fluid (as indicated by the arrows)towards the target separation material 3402, can be used to reduce atleast some of the coating off of the separation material. This cancreate a preferential change in the amount of coating in selected areasof the separation material. One example may show removal or at leastreduction of coating on one side of the separation material. Optionally,some may direct the wash fluid to wash coating off of an interior regionof the separation material. Other configurations where portions ofcoating are removed from other select areas are not excluded.

In one embodiment described herein, a carefully controlled washing isdesirable so as to not completely remove the hemolysis preventingagent—which would result in hemolysis. In contrast, insufficient washwill result in sufficient amount of the hemolysis preventing agentleaching into the plasma and causing hemolysis. Thus, in onenon-limiting example, a reduced amount of coating, or coating ininterior portions of the separation material can be acceptable.Optionally, as seen in FIG. 82, some embodiments may also use a bath3410 of wash fluid that preferentially removes coating material fromcertain areas of the separation material. Optionally, spray washing andbath soaking, or vice versa, may be combined for use on a separationmaterial. This processing may occur sequentially or simultaneously.

2. Custom separation material coating: In another embodiment describedherein, both coated and uncoated versions of the plasma separationmaterial can be coated using a custom formulation which is compatiblewith assay chemistries. The coating may contain one or more of thefollowing: proteins, surfactants, sugars, organic and inorganic salts,anti-coagulants, etc. In one non-limiting example, the coating could beapplied to an initially un-coated separation material to preventhemolysis. Optionally, an initially coated separation material may befurther coated to prevent assay interfering substances from leachinginto the bodily fluid from the separation material.

3. Charge Neutralization: In one embodiment described herein, separationmaterial surface charge can be neutralized to prevent retention ofsmall, oppositely charged ions. For example, the separation materialwith NTA coating has a negatively-charged surface, which can beneutralized to prevent retention of positively charged Ca++ ions.Optionally, if a coating has a positively-charged surface and is in turnattracting negatively charged ions in a detrimental manner, the memberwill be treated to neutralize the undesired charge condition.

4. Other techniques and/or materials may also be used to create a filtersuch as a separation material that has anti-hemolytic qualities on thecapture surfaces of the filter and non-leaching qualities on othersurfaces of the filter. Some embodiments may combine one or more of theforegoing techniques on a separation material. By way of non-limitingexample, one embodiment may have coated and uncoated regions on aseparation material along with having been treated to achieve chargeneutralization before, during, and/or after coating.

EXAMPLES

Using a dynamic wash technique, asymmetric membranes were washed withhigh performance liquid chromatography (HPLC) grade water and thentested. In one non-limiting example, the membrane has a pore volume of24 per 10 mm² of membrane. The pore loading is defined as the ratio ofthe total volume of blood to the pore volume. For a blood volume of 404with membrane surface area of 100 mm², this corresponds to a poreloading of 2×. The wash procedure comprised pre-mounting membrane in afixture for filtration. In this particular example, about 600 uL ofwater is directed through the membrane and then the water is discarded.This wash process of directing water through the membrane was repeated,which in this particular example, involved repeating the wash five (5)times. After washing, the membranes are allowed to dry. Filtration ofthe dynamically washed membranes were then tested.

Washing by way of soaking (“static wash”) rather than the flow-throughtechnique (“dynamic wash”) can create differences in the performance ofthe resulting membrane. In at least some static washed membranes,anti-hemolytic is preferentially removed from the large pore region. Inat least some dynamic wash membranes, anti-hemolytic is preferentiallyremoved from the small pore region. This asymmetry in coating materialmay be desirable when the formed blood components contact the membranewhere the pores are larger while only plasma contacts the smallestpores. Hemolysis prevention happens only in the regions where RBCs canenter or be contacted (i.e. the large pore region). It is not possibleto hemolyze plasma and thus coating the small pore region withanti-hemolytic does not result in noticeable performance benefit. Asnoted herein, the excess anti-hemolytic may have adverse impact on assayresults for the assays sensitive to excess anti-hemolytic coating.

In static wash, diffusion dominates removal of anti-hemolytic. In someembodiments of the membrane, large pores may be ˜50× bigger than smallpores. Mass diffusion rate is proportional to cross sectional flow area.Thus diffusion rate of anti-hemolytic away from membrane on large poreside may be ˜2500× greater than on small pore side. Thus, without beingbound to any particular theory, total removal should be much greater onlarge pore side, where the RBCs contact the membrane.

In dynamic wash, shear dominates removal of anti-hemolytic. Shearincreases dramatically with decreasing diameter. Without being bound toany particular theory, total removal should be greater in small poreregions, where shear is most significant.

In yet another embodiment, the coating on the membrane can be a materialthat provides a negative charge. Without being bound to any particulartheory, a negative charge repels formed blood component that have anegative polarity, and thereby reduces mechanical trauma inflicted onsuch formed blood components via contact with the membrane duringfiltration. Some embodiments may use formulations with negativelycharged substances to coat all or optionally selective areas on themembrane. One embodiment may use casein 0.5%, Tween 20 1.35%, sucrose5%, 15 minute soak time. Optionally, one embodiment may use Li-Heparin50mg/mL, sucrose 5%. Optionally, one embodiment may use Li-Heparin50mg/mL, Tween 80 1.35%, sucrose 5%. Optionally, one embodiment may useCasein 1.0%, Tween 20 2.70%, sucrose 5%. Optionally, one embodiment mayuse Li-Heparin 100 mg/mL, Tween 20 2.70%, sucrose 5%.

Sample Processing

Referring now to FIG. 83, one embodiment of bodily fluid samplecollection and transport system will now be described. FIG. 83 shows abodily fluid sample B on a skin surface S of the subject. In thenon-limiting example of FIG. 83, the bodily fluid sample B can becollected by one of a variety of devices. By way of non-limitingexample, collection device 1530 may be but is not limited to thosedescribed herein. In the present embodiment, the bodily fluid sample Bis collected by one or more capillary channels and then directed intosample vessels 1540. The sample B forms through a wound that may beformed on the subject. This may be by way of fingerstick or woundcreated at other alternate sites on the body. By way of non-limitingexample, a lancet, a needle, other penetrating device, or othertechnique may be used to release the bodily fluid sample from thesubject. By way of non-limiting example, at least one of the samplevessels 1540 may have an interior that is initially under a partialvacuum that is used to draw bodily fluid sample into the sample vessel1540. Some embodiments may simultaneously draw sample from the samplecollection device into the sample vessels 1540 from the same ordifferent collection channels in the sample collection device.Optionally, some embodiments may simultaneous draw sample into thesample vessels.

In the present embodiment after the bodily fluid sample is inside thesample vessels 1540, the sample vessels 1540 in their holder 1542 (oroptionally, removed from their holder 1542) may placed in the sampleverification device or directly into a storage device in a temperaturecontrolled environment. In the present embodiment after the sampleverification is completed, the sample vessels 1540 in their holder 1542(or optionally, removed from their holder 1542) are loaded into thetransport container 1500. In one non-limiting example, one of the samplevessels 1540 may contain only liquid portions of the sample (no formedblood components) which the other may contain sample with both liquidportion and formed component portion. In another non-limiting example,at least two of the sample vessels 1540 may contain only liquid portionsof the sample (no formed blood components).

In this embodiment, there may be one or more slots sized for the samplevessel holder 1542 or slots for the sample vessels in the transportcontainer 1500. By way of non-limiting example, they may hold the samplevessels in an arrayed configuration and oriented to be vertical or someother pre-determined orientation. It should be understood that someembodiments of the sample vessels 1540 are configured so that they holddifferent amount of sample in each of the vessels. By way ofnon-limiting example, this can be controlled based on the amount ofvacuum force in each of the sample vessels, the amount of samplecollected in the sample collection channel(s) of the collection device,and/or other factors. Optionally, different pre-treatments such as butnot limited to different anti-coagulants or the like can also be presentin the sample vessels.

As seen in FIG. 83, the sample vessels 1540 are collecting sample at afirst location such as but not limited to a sample collection site. Byway of non-limiting example, the bodily fluid samples are thentransported in the transport container 1500 to a second location such asbut not limited to an analysis site. The method of transport may be bycourier, postal delivery, or other shipping technique. In manyembodiments, the transport may be implemented by having a yet anothercontainer that holds the transport container therein. In one embodiment,the sample collection site may be a point-of-care. Optionally, thesample collection site is a point-of-service. Optionally, the samplecollection site is remote from the sample analysis site.

Although the present embodiment of FIG. 83 shows the collection ofbodily fluid sample from a surface of the subject, other alternativeembodiments may use collection techniques for collecting sample fromother areas of the subject, such as by venipuncture, to fill the samplevessel(s) 1540. Such other collection techniques are not excluded foruse as alternative to or in conjunction with surface collection. Surfacecollection may be on exterior surfaces of the subject. Optionally, someembodiments may collect from accessible surfaces on the interior of thesubject. Presence of bodily fluid sample B on these surfaces may benaturally occurring or may occur through wound creation or othertechniques to make the bodily fluid surface accessible.

Referring now to FIGS. 84 to 99, still other embodiments of manifoldsand distribution patterns of the channels to the separator will now bedescribed. In one embodiment, the device is configured for transversefilling of the channels along a shorter planar dimension of theseparator versus along a lengthwise filling direction along a longdimension of the separator, particularly in a separator with an aspectratio where at least one dimension along one axis is shorter thananother dimension along another axis in the same plane.

In one embodiment as seen in the cross-sectional view of FIG. 84,whether the plenum 4000 is separate from the whole blood channel or not,the lead-ins will be angled through channel(s) 4002 that route bloodflow to distribution channel(s) 4004 over a separator 4006, such as butnot limited to the membrane. Optionally, some embodiments may include avent 4008 at one end or other position along the distribution channel4004. As seen in this embodiment in FIG. 84, the channel(s) 4002 fromthe plenum may be angled (from above or below) and not horizontal likethe distribution channel 4004. By way of non-limiting example, theangled conduit form plenum to distribution channels and membrane is onlyone embodiment of how sample may be transported from the plenum.Optionally, some embodiments may have both the channel 4002 and thedistribution channel(s) 4004 parallel to each other and in the sameplane. As seen in FIG. 84, there may be a narrow area 4010 along thetransition between the channel 4002 and the distribution channel(s)4004. A perspective view may be found in FIG. 27D.

As seen in FIGS. 84 and 27D, an upper portion of sample in plenum 4000is drawn into the channels 4002 to be moved along a path to separationsuch as into plasma when the sample is blood. A portion of sample thatremains in the plenum 4000 travels along a different path and iscollected without going through a separator. As seen in FIG. 27D, onpath exits via arrow 4022 (separated plasma) and the other pathway exitsvia arrow 4024 (whole blood).

FIGS. 85A-85D show various filling patterns. Illustrations of differentfilling scenarios in the manifold is shown with combined plenum. FIG.85A: Flow is essentially even and parallel. Sometimes there is a lagbetween the completion of plenum filling, and initiation of the lead-inflow. FIG. 85B: Filling of the distribution channels and membrane occursfrom upstream end to downstream. Based on previous observations, thistype of behavior frequently corresponds to a situation where thelead-ins direct flow to the membrane as the plenum fills, rather thanafter the plenum filling has completed. FIG. 85C: Filling order is fromdownstream to upstream, which is the reverse of what is shown in FIG.85B. FIG. 85(d): One group of channels lags the others.

FIG. 86 shows one embodiment where the plenum runs over the top of theseparator and not through it or to the side. The membrane may becompressed to seal around the plenum 4030. A separate channel 4020 inthe device may be used for whole blood that will not be separated intoplasma. Unlike the embodiment of FIG. 84 where a single channel extendsfrom an entrance to an exit containing both sample that will beseparated and sample that will exit without being separated, thisembodiment of FIG. 86 uses separate channels 4020 and 4030 for thesamples, depending on how the sample will be processed.

FIGS. 87 and 88 show embodiments where additional membrane compressionis employed around the perimeter of the plenum 4030 (except in theregions where that compression cannot be achieved due to the presence ofthe lead-ins). In this non-limiting example, this creates a membranecompression zone 4050.

Referring now to the embodiment of FIG. 89, one modification if theuncompressed membrane under the lead-ins is problematic is to use a thinfilm 4040 placed onto the bottom of the plenum area to create a “floor”for this channel, which separates the blood entirely from thenon-functional region of the membrane underneath. The film may be formedfrom an inert material that does not interact with sample in the plenum4030. Optionally, the film may be Teflon film, a polymer film, a plasticfilm, or other material configured to not impact sample quality.

Referring now to FIG. 90, an additional variation of the devicecomprises placing the plenum on top of the membrane. In this variationthe distribution channels do not branch, but rather they all connectdirectly to the plenum. Because angled through channels are not usedwhen the plenum 4030 is directly on top of the membrane, it is notnecessary to minimize the number of lead-ins by means of branching:there are no small, angled core pins or flash to worry about. Providedthat the plenum region of the membrane can be adequately decoupled fromthe functional region of the membrane, each distribution channel 4036could be directly fed by the plenum 4030. In this non-limiting example,the benefit of this configuration is that it would enable betteruniformity in the distribution of the blood over the membrane, andbetter control of the distribution of blood over the membrane.

Referring now to FIG. 91, the configurations with the plenum separatefrom the membrane use angled through channels for their lead-ins. Thisis noted in FIG. 91. In this embodiment, the whole blood to exit fromexit 4024 is drawn from the same plenum 4000 as sample on a path to exita separate sample from exit 4022. FIG. 91 shows that in some embodimentsa common channel is used to provide sample to an outlet for outputtingunseparated sample through outlet 4024 and for directing a portion ofthe sample along a path to a second outlet 4022. FIG. 91 shows that thepathway through the separator may be by way of a single channel 4002(angled or not) from the common channel and that this single channel maythen spread to form multiple channels 4004 over the separation material.The transition from a single channel 4002 to multiple channels 4004 maymore evenly distribute the sample. As seen in FIG. 91, the channels 4004may have different shapes depending on the desired distribution pattern.FIG. 91 shows that channels 4004 may form a “fork” pattern as theyextend away from the channel 4002. Some embodiments may form otherpatterns such as curved, spiral, star, radial tire spoke, or othergeometric pattern. Most of these patterns are in direction lateral to,orthogonal to, or along the short axis, relative to a long longitudinalaxis of the separation material.

Referring now to FIGS. 92 and 93, there are two layouts shown with theplenum on the side nearest the whole blood channel. FIG. 92 shows aversion in which the plenum is centered on the inlet. There areadvantages to the in-line configuration, such as simplicity and the factthat this layout will probably encourage rapid filling of the membrane.FIG. 94 shows a still further perspective view of the

FIG. 93 shows yet another embodiment with the plenum adjacent to thewhole blood channel. Here is that the plenum 4030 is not directly inline with the inlet. Rather, there is a curve just downstream of theintersection between the plenum and the inlet. What is gained is morerange in the membrane aspect ratios that fit on the device.

FIGS. 95A to 95D show various embodiments of distribution channelvariation, from large and widely spaced, to no channels at all. As seenin those FIGS. 95A to 95D, there may be a vent 4008 at the end of thevarious distribution channels.

FIGS. 96 and 97 show two possible configurations that employdeliberately unequal distribution of the blood over a separator such asbut not limited to the membrane. FIG. 96 shows that the pitch betweendistribution channels 4036 can be reduced at one of end of the device.FIG. 97 shows that in addition to pitch variation between distributionchannels, some embodiments may also various the diameter orcross-sectional area so that some channels have reduced size closer towhere the sample is entering the plenum, so that more is directedtowards those channels located further away from the inlet of theplenum. This may more preferentially direct sample flow so that thedistribution of the sample is more even, without an initial rush thatmainly fills the channels closest to the inlet of the plenum as seen inFIG. 85B. Optionally, some embodiments may combine the features of FIGS.96 and 97.

Referring now to FIG. 98, in one embodiment, the bed of capillarychannels at the back of the membrane serves the dual purposes of helpingthe plasma to wick through the membrane, and providing a low-resistanceflow pathway external to the membrane for the plasma to flow throughduring the extraction process. FIG. 98 shows perspective view of a“lower” portion of the device that is typically on the underside of themembrane, showing those structures that handle the fluid that has passedthrough the separation membrane. Optionally, capillary surfaces may beconfigured to exhibit the behavior known as “total wetting” in thepresence of plasma. This corresponds to a contact angle of zero, meaningthat the liquid will continue to spread until the volume of thecapillaries is filled with plasma. This helps to achieve even flowthrough the membrane both prior to and during the extraction process. Asseen in the magnified view of FIG. 98, at one end of the capillarychannels near the extraction port to exit 4022, the channels extend toan area just short of the end, so that a channel area 4060 is formed toroute separated sample such as but not limited to plasms to theextraction port.

FIG. 99 shows one embodiment where the membrane is compressed atlocation 4100 to provide a sealed perimeter. In one embodiment, thethickness used for compression is 100 μm. In one embodiment, thethickness used for compression is at least 100 μm. In one embodiment,the thickness used for compression is at least 90 μm but less than 110μm. In one embodiment, the thickness used for compression is at least 80μm but less than 110 μm. In one embodiment, the thickness used forcompression is at least 70 μm but less than 110 μm. In one embodiment,the thickness used for compression is at least 60 μm but less than 110μm. In one embodiment, the thickness used for compression is at least 50μm but less than 110 μm.

While the teachings has been described and illustrated with reference tocertain particular embodiments thereof, those skilled in the art willappreciate that various adaptations, changes, modifications,substitutions, deletions, or additions of procedures and protocols maybe made without departing from the spirit and scope of the invention.For example, with any of the above embodiments, it should be understoodthat the fluid sample may be whole blood, diluted blood, interstitialfluid, sample collected directly from the patient, sample that is on asurface, sample after some pre-treatment, or the like. Although theembodiments herein are described in the context of an anti-hemolyticcoating, it should be understood that these embodiments may also beconfigured for use with other types of coatings, including but notlimited to other coatings which may undesirably mix into the bodilyfluid upon prolonged fluid exposure. Other material used withembodiments herein may include but is not limited to one or more of thefollowing: anti-coagulants, proteins (BSA, HSA, Heparin, Casein, etc.),surfactants (Tween, Silwet, SDS, etc.), sugars (sucrose, trealose,etc.). It should be understood that in some embodiments, coatings of oneor more of the following may be used to coat portions of fluid pathwaysof the device, only the channels of the distributor, only thenon-channel portions of the distributor, sample collection areas, sampledistributor area(s), channels, tubes, chambers, or other features of thedevice with: anti-hemolytic, anti-coagulants, proteins (BSA, HSA,Heparin, Casein, etc.), surfactants (Tween, Silwet, SDS, etc.), sugars(sucrose, trealose, etc.), or other coatings.

Although the embodiments herein are described in the context ofcapturing formed components such as blood cells or platelets, it shouldbe understood that these embodiments can also be adapted for use withfluid containing other solid, semi-solid, or formed components orparticles. Although the embodiments herein are described in the contextof separation material, it should be understood that these embodimentscan also be adapted for use other filter materials such as meshes,porous layers, or other layer like materials or structures.

In one embodiment described herein, a bodily fluid separation materialis provided comprising a formed component capture region and a bodilyfluid pass-through region. The pass-through region has structures with areduced liquid leaching quality relative to than the capture region,wherein during separation material use, bodily fluid enters the captureregion prior to entering the pass-through region. Optionally, a bodilyfluid pass-through region has a reduced amount of liquid leachingmaterial relative to than the capture region.

In another embodiment described herein, a bodily fluid separationmaterial is provided comprising an anti-hemolytic and formed componentcapture region; and a bodily fluid pass-through region having lessanti-hemolytic material than the capture region, wherein duringseparation material use, bodily fluid enters the capture region prior toentering the pass-through region.

In yet embodiment described herein, a bodily fluid separation materialis provided comprising a first filter region of the separation materialhaving an anti-hemolytic coating and mesh spacing sized to constrainformed blood components therein; a second filter region of theseparation material having mesh spacing smaller than mesh spacing of thefirst filter region and configured to have an amount of anti-hemolyticcoating less than that of the first region.

In a still further embodiment described herein, a bodily fluidseparation material is provided comprising a percolating network ofstructures wherein a first region of the percolating network with ananti-hemolytic coating on structures in the region, said structuressized and spaced to allow formed blood components to enter the firstregion but constraining blood components therein from passing completelythrough the first region; and a second region of the percolating networkwith a reduced anti-hemolytic coating on structures sized and spaced toprevent formed blood components from entering the second region, whereinbodily fluid passes through the first region prior to reaching thesecond region.

It should be understood that embodiments herein may be adapted toinclude one or more of the following features. For example, theseparation material may be an asymmetric separation material.Optionally, the anti-hemolytic material on the separation materialcomprises single and/or double alkyl chain N-oxides of tertiary amines(NTA). Optionally, the first region comprises a first separationmaterial layer and the second region comprises a second separationmaterial layer. Optionally, the separation material comprises a firstseparation material coupled to a second separation material. Optionally,the separation material comprises at least two separate separationmaterials. Optionally, there may be at least another region of theseparation material between the first region and the second region.Optionally, the first region of the separation material may be in fluidcommunication with the second region. Optionally, the first region maybe spaced apart from the second region.

In yet another embodiment described herein, a method is provided forforming a bodily fluid separation material. The method comprises coatingthe separation material with an anti-hemolytic coating on a first regionand a second region of the separation material; reducing anti-hemolyticeffect of the second region of the separation material relative to thefirst region, wherein when the separation material is in operation,bodily fluid passes through the first region prior to reaching thesecond region.

It should be understood that embodiments herein may be adapted toinclude one or more of the following features. For example, the methodmay include reducing the anti-hemolytic effect by washing off at least aportion of the anti-hemolytic coating on the second region. Optionally,washing off comprises directing solvent through the separation material.Optionally, washing off comprises soaking only a portion of theseparation material in a solvent. Optionally, reducing theanti-hemolytic effect comprises adding another coating of a differentmaterial over the anti-hemolytic coating on the second region.Optionally, reducing the anti-hemolytic effect comprises treating theseparation material to bring its electrical charge state to a neutralstate and thus reduce the attraction of ions that increase theanti-hemolytic effect.

In yet another embodiment described herein, a method is provided forforming a bodily fluid separation material. The method comprises coatingat least a first region of the separation material with ananti-hemolytic coating; not coating at least second region of theseparation material with the anti-hemolytic coating. Optionally, someembodiments have a bilayer structure based on a substantially evencoating of anti-hemolytic material, but instead has a region ofsubstantially greater pore size than another region. Although thematerial may be asymmetric, it is a not a linear gradient, but insteadhas a rapid change in pore size at an inflection point when pore size isgraphed in depth from top of the layer to bottom of the layer.

Additionally, concentrations, amounts, and other numerical data may bepresented herein in a range format. It is to be understood that suchrange format is used merely for convenience and brevity and should beinterpreted flexibly to include not only the numerical values explicitlyrecited as the limits of the range, but also to include all theindividual numerical values or sub-ranges encompassed within that rangeas if each numerical value and sub-range is explicitly recited. Forexample, a size range of about 1 nm to about 200 nm should beinterpreted to include not only the explicitly recited limits of about 1nm and about 200 nm, but also to include individual sizes such as 2 nm,3 nm, 4 nm, and sub-ranges such as 10 nm to 50 nm, 20 nm to 100 nm, etc. . . .

The publications discussed or cited herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.All publications mentioned herein are incorporated herein by referenceto disclose and describe the structures and/or methods in connectionwith which the publications are cited. The following applications arefully incorporated herein by reference for all purposes: U.S. Pat. App.Ser. No. 62/051,929, filed Sep. 17, 2014; U.S. Pat. No. 8,088,593; U.S.Pat. No. 8,380,541; U.S. patent application Ser. No. 13/769,798, filedFeb. 18, 2013; U.S. patent application Ser. No. 13/769,779, filed Feb.18, 2013; U.S. Pat. App. Ser. No. 61/766,113 filed Feb. 18, 2013, U.S.patent application Ser. No. 13/244,947 filed Sep. 26, 2011;PCT/US2012/57155, filed Sep. 25, 2012; U.S. application Ser. No.13/244,946, filed Sep. 26, 2011; U.S. patent application Ser. No.13/244,949, filed Sep. 26, 2011; U.S. application Ser. No. 61/673,245,filed Sep. 26, 2011; U.S. Patent Application Ser. No. 61/786,351 filedMar. 15, 2013; U.S. Patent Application Ser. No. 61/948,542 filed Mar. 5,2014; U.S. Patent Application Ser. No. 61/952,112 filed Mar. 12, 2014;U.S. Patent Application Ser. No. 61/799,221 filed Mar. 15, 2013, U.S.Patent Application Ser. No. 61/697,797 filed Sep. 6, 2012, U.S.Provisional Patent Application, 62/216,359 filed Sep. 9, 2016, and U.S.Patent Application Ser. No. 61/733,886 filed Dec. 5, 2012, thedisclosures of which patents and patent applications are all herebyincorporated by reference in their entireties for all purposes.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. Any feature, whetherpreferred or not, may be combined with any other feature, whetherpreferred or not. The appended claims are not to be interpreted asincluding means-plus-function limitations, unless such a limitation isexplicitly recited in a given claim using the phrase “means for.” Itshould be understood that as used in the description herein andthroughout the claims that follow, the meaning of “a,” “an,” and “the”includes plural reference unless the context clearly dictates otherwise.For example, a reference to “an assay” may refer to a single assay ormultiple assays. Also, as used in the description herein and throughoutthe claims that follow, the meaning of “in” includes “in” and “on”unless the context clearly dictates otherwise. Finally, as used in thedescription herein and throughout the claims that follow, the meaning of“or” includes both the conjunctive and disjunctive unless the contextexpressly dictates otherwise. Thus, the term “or” includes “and/or”unless the context expressly dictates otherwise.

1-29. (canceled)
 29. A sample collection device, comprising: (a) a body,containing a collection channel , the collection channel comprising afirst opening and a second opening, and being configured to draw abodily fluid via capillary action from the first opening towards thesecond opening; (b) a base, containing a sample container for receivingthe bodily fluid sample, the sample container being engagable with thecollection channel, having an interior with a vacuum therein, and havinga cap configured to receive a channel; and (c) a support, wherein, thebody and the base are connected to opposite ends of the support, and areconfigured to be movable relative to each other, such that samplecollection device is configured to have an extended state and acompressed state, wherein at least a portion of the base is closer tothe body in the extended state of the device than in the compressedstate, the second opening of the collection channel is configured topenetrate the cap of the sample container, in the extended state of thedevice, the second opening of the collection channel is not in contactwith the interior of the sample container, and in the compressed stateof the device, the second opening of the collection channel extends intothe interior of the sample container through the cap of the container,thereby providing fluidic communication between the collection channeland the sample container; a separator along the sample collectionchannel, the separator configured to remove formed component from thesample prior to and when outputting to the sample container.
 30. Asample collection device, comprising: (a) a body, containing acollection channel , the collection channel comprising a first openingand a second opening, and being configured to draw a bodily fluid viacapillary action from the first opening towards the second opening; (b)a base, containing a sample container for receiving the bodily fluidsample, the sample container being engagable with the collectionchannel, having an interior with a vacuum therein and having a capconfigured to receive a channel; (c) a support, and (d) an adaptorchannel, having a first opening and a second opening, the first openingbeing configured to contact the second opening of the collectionchannel, and the second opening being configured to penetrate the cap ofthe sample container, wherein, the body and the base are connected toopposite ends of the support, and are configured to be movable relativeto each other, such that sample collection device is configured to havean extended state and a compressed state, wherein at least a portion ofthe base is closer to the body in the extended state of the device thanin the compressed state, in the extended state of the device, theadaptor channel is not in contact with one or both of the collectionchannel and the interior of the sample container, and in the compressedstate of the device, the first opening of the adaptor channel is incontact with the second opening of the collection channel, and thesecond opening of the adaptor channel extends into the interior of thesample container through the cap of the container, thereby providingfluidic communication between the collection channel and the samplecontainer; a separator along the sample collection channel, theseparator configured to remove formed component from the sample prior toand when outputting to the sample container.
 31. (canceled) 32.(canceled)
 33. (canceled)
 34. (canceled)
 35. A method comprising:collecting a bodily fluid sample into device having a first collectionchannel and a second collection channel, the collection channelcomprising a first opening and a second opening, and being configured todraw the bodily fluid via capillary action from the first openingtowards the second opening; and using a separator along the first samplecollection channel to remove formed component from the sample prior toand when outputting to the sample container; wherein when the bodilyfluid sample is blood, the device outputs both blood and plasma, eachfrom separate outlets, from the one sample collected into the device.36-46. (canceled)
 47. The device of claim 29 further comprising a fillindicator to indicate when a minimum fill level has been reached. 48.The device of claim 29 wherein the separator comprises separationmaterial held in the device under compression.
 49. The device of claim29 wherein the separator comprises an asymmetric porous membrane. 50.The device of claim 29 wherein the separator comprises a mesh.
 51. Thedevice of claim 29 wherein at least a portion of the separator comprisesa polyethersulfone.
 52. The device of claim 29 wherein at least aportion of the separator comprises an asymmetric polyethersulfone. 53.The device of claim 30 wherein at least a portion of the separatorcomprises polyarylethersulfone.
 54. The device of claim 30 wherein atleast a portion of the separator comprises an asymmetricpolyarylethersulfone.
 55. The device of claim 30 wherein at least aportion of separator comprises a polysulfone.
 56. The device of claim 30wherein the separator comprises an asymmetric polysulfone.
 57. Thedevice of claim 30 wherein the separator comprises a polysulfonemembrane.